Compounds for inflammation and immune-related uses

ABSTRACT

The invention relates to compounds of structural formulas (I), (VII) and (XI): 
     
       
         
         
             
             
         
       
     
     or a pharmaceutically acceptable salt, solvate, clathrate, or prodrug thereof, wherein X 1 , X 2 , X 3 , Y, Z, L, R 1 , R 2 , R 3 , R 18  and n are defined herein. These compounds are useful as immunosuppressive agents and for treating and preventing inflammatory conditions, allergic disorders, and immune disorders.

RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional PatentApplication No. 60/642,179, filed on Jan. 7, 2005 and U.S. ProvisionalPatent Application No. 60/707,845, filed on Aug. 12, 2005. The entireteachings of each of these applications are incorporated herein byreference.

FIELD OF THE INVENTION

This invention relates to biologically active chemical compounds, namelybiphenyl and pyridylphenyl derivatives that may be used forimmunosuppression or to treat or prevent inflammatory conditions, immunedisorders, and allergic disorders.

BACKGROUND OF THE INVENTION

Inflammation is a mechanism that protects mammals from invadingpathogens. However, while transient inflammation is necessary to protecta mammal from infection, uncontrolled inflammation causes tissue damageand is the underlying cause of many illnesses. Inflammation is typicallyinitiated by binding of an antigen to T-cell antigen receptor. Antigenbinding by a T-cell initiates calcium influx into the cell via calciumion channels, such as Ca²⁺-release-activated Ca²⁺ channels (CRAC).Calcium ion influx in turn initiates a signaling cascade that leads toactivation of these cells and an inflammatory response characterized bycytokine production.

Interleukin 2 (IL-2) is a cytokine that is secreted by T cells inresponse to calcium ion influx into the cell. IL-2 modulatesimmunological effects on many cells of the immune system. For example,it is a potent T cell mitogen that is required for the T cellproliferation, promoting their progression from G1 to S phase of thecell cycle; it stimulates the growth of NK cells; and it acts as agrowth factor to B cells and stimulates antibody synthesis.

IL-2, although useful in the immune response, can cause a variety ofproblems. IL-2 damages the blood-brain barrier and the endothelium ofbrain vessels. These effects may be the underlying causes ofneuropsychiatric side effects observed under IL-2 therapy, e.g. fatigue,disorientation and depression. It also alters the electrophysiologicalbehaviour of neurons.

Due to its effects on both T and B cells, IL-2 is a major centralregulator of immune responses. It plays a role in inflammatoryreactions, tumour surveillance, and hematopoiesis. It also affects theproduction of other cytokines, inducing IL-1, TNF-α and TNF-β secretion,as well as stimulating the synthesis of IFN-γ in peripheral leukocytes.

T cells that are unable to produce IL-2 become inactive (anergic). Thisrenders them potentially inert to any antigenic stimulation they mightreceive in the future. As a result, agents which inhibit IL-2 productioncan be used for immunosupression or to treat or prevent inflammation andimmune disorders. This approach has been clinically validated withimmunosuppressive drugs such as cyclosporin, FK506, and RS61443. Despitethis proof of concept, agents that inhibit IL-2 production remain farfrom ideal. Among other problems, efficacy limitations and unwanted sideeffects (including dose-dependant nephrotoxicity and hypertension)hinder their use.

Over production of proinflammatory cytokines other than IL-2 has alsobeen implicated in many autoimmune diseases. For example, Interleukin 5(IL-5), a cytokine that increases the production of eosinophils, isincreased in asthma. Overproduction of IL-5 is associated withaccumulation of eosinophils in the asthmatic bronchial mucosa, a hallmark of allergic inflammation. Thus, patients with asthma and otherinflammatory disorders involving the accumulation of eosinophils wouldbenefit from the development of new drugs that inhibit the production ofIL-5.

Interleukin 4 (IL-4) and interleukin 13 (IL-13) have been identified asmediators of the hypercontractility of smooth muscle found ininflammatory bowel disease and asthma. Thus, patients with asthma andinflammatory bowel disease would benefit from the development of newdrugs that inhibit IL-4 and IL-13 production.

Granulocyte macrophage-colony stimulating factor (GM-CSF) is a regulatorof maturation of granulocyte and macrophage lineage population and hasbeen implicated as a key factor in inflammatory and autoimmune diseases.Anti-GM-CSF antibody blockade has been shown to ameliorate autoimmunedisease. Thus, development of new drugs that inhibit the production ofGM-CSF would be beneficial to patients with an inflammatory orautoimmune disease.

There is therefore a continuing need for new drugs which overcome one ormore of the shortcomings of drugs currently used for immunosuppressionor in the treatment or prevention of inflammatory disorders, allergicdisorders and autoimmune disorders.

Desirable properties of new drugs include efficacy against diseases ordisorders that are currently untreatable or poorly treatable, newmechanism of action, oral bioavailability and/or reduced side effects.

SUMMARY OF THE INVENTION

This invention meets the above-mentioned needs by providing certainbiphenyl and phenylpyridyl derivatives that inhibit the activity of CRAGion channels and inhibit the production of IL-2, IL-4, IL-5, IL-13,GM-CSF, TNF-α, and IFNγ. These compounds are particularly useful forimmunosuppression and/or to treat or prevent inflammatory conditions andimmune disorders.

One embodiment of the invention relates to compounds of formula (I):

-   -   or a pharmaceutically acceptable salt, solvate, clathrate, or        prodrug thereof, wherein:    -   L is a linker selected from the group consisting of a covalent        bond, —NRCH₂—, —CH₂NR—, —C(O)—, —NR—C(O)—, —C(O)—NR—, —OC(O)—,        —C(O)O—, —C(S)—, —NR—C(S)—, —C(S)—NR—;    -   X₁ and X₃ are each, independently, CH or N;    -   X₂ is CH, CR₁₀ or N;    -   each Z is independently selected from the group consisting of a        lower alkyl, a lower haloalkyl, a halo, a lower alkoxy, a lower        alkyl sulfanyl, cyano, nitro, or lower haloalkoxy;    -   R, for each occurrence is independently selected from —H, an        alkyl, —C(O)R₅, or —C(O)OR₅;    -   R₁ and R₂ are each, independently, a halo, a haloalkyl, a lower        alkyl, a lower alkoxy, or a haloalkoxy;    -   R₃ is an alkyl, a haloalkyl, a halo, a haloalkoxy, —OR₅, —SR₅,        or —NR₆R₇;    -   R₁₈ is a halo, cyano, nitro, —C(O)R₅, —C(O)OR₅, —C(O)SR₅,        —C(O)NR₆R₇, —C(S)R₅, —C(S)OR₅, —C(S)SR₅, —C(S)NR₆R₇, —C(NR₈)R₅,        —C(NR₈)OR₅, —C(NR₈)SR₅, —C(NR₈)NR₈R₇, —S(O)_(p)R₅, —S(O)_(p)NR₅,        —S(O)_(p)OR₅, —P(O)(OR₅)₂, —OP(O)(OR₅)₂, —P(O)(R₅)₂, a five or        six membered optionally substituted heterocycloalkyl, a five or        six membered optionally substituted heterocyclyl, or a five or        six membered optionally substituted heteroaryl;    -   R₅, for each occurrence, is independently, H, an optionally        substituted alkyl, an optionally substituted alkenyl, an        optionally substituted alkynyl, an optionally substituted        cycloalkyl, an optionally substituted cycloalkenyl, an        optionally substituted heterocyclyl, an optionally substituted        aryl, an optionally substituted heteroaryl, an optionally        substituted aralkyl, or an optionally substituted heteraralkyl;    -   R₆ and R₇, for each occurrence are, independently, H, an        optionally substituted alkyl, an optionally substituted alkenyl,        an optionally substituted alkynyl, an optionally substituted        cycloalkyl, an optionally substituted cycloalkenyl, an        optionally substituted heterocyclyl, an optionally substituted        aryl, an optionally substituted heteroaryl, an optionally        substituted aralkyl, or an optionally substituted heteraralkyl;        or R₆ and R₇ taken together with the nitrogen to which they are        attached are an optionally substituted heterocyclyl or        optionally substituted heteroaryl;    -   R₈, for each occurrence, is independently —H, a halo, an alkyl,        —OR₅, —NR₆R₇, —C(O)R₅, —C(O)OR₅, or —C(O)NR₆R₇;    -   R₁₀ is a lower alkyl, a lower alkoxy, a halo, a lower haloalkyl,        a lower haloalkoxy, a cyano, nitro, —C(O)R₅, —C(O)OR₅, —C(O)SR₅,        —C(O)NR₆R₇, —C(S)R₅, —C(S)OR₅, —C(S)SR₅, —C(S)NR₆R₇, —C(NR₈)R₅,        —C(NR₈)OR₅, —C(NR₈)SR₅, —C(NR₈)NR₆R₇, —S(O)_(p)R₅, —P(O)(OR₅)₂,        —OP(O)(OR₅)₂, or —P(O)(R₅)₂;    -   n is zero or an integer from 1 to 4; and    -   p, for each occurrence, is independently 1 or 2.

In one embodiment, the invention relates to compounds of formula (II):

-   -   or a pharmaceutically acceptable salt, solvate, clathrate, or        prodrug thereof, wherein:    -   L, X₁, X₂, X₃, Z, R₁, R₂, R₃ and n are defined as for formula        (I); and    -   R₄ is a halo, cyano, nitro, —C(O)R₅, —C(O)OR₅, —C(O)SR₅,        —C(O)NR₆R₇, —C(S)R₅, —C(S)OR₅, —C(S)SR₅, —C(S)NR₆R₇, —C(NR₈)R₅,        —C(NR₈)OR₅, —C(NR₈)SR₅, —C(NR₈)NR₆R₇, —S(O)_(p)R₅, —S(O)_(p)NR₅,        —S(O)_(p)OR₅, —P(O)(OR₅)₂, —OP(O)(OR₅)₂, —P(O)(R₅)₂, or an        ester, amide or carboxylic acid bioisostere.

In another embodiment, the invention relates to compounds of formula(VII):

-   -   or a pharmaceutically acceptable salt, solvate, clathrate, or        prodrug thereof, wherein:    -   X₁, L, Z, R₁, R₃, R₁₈ and n are defined as for formula (I).

In another embodiment, the invention relates to compounds represented byformula (VIII):

-   -   or a pharmaceutically acceptable salt, solvate, clathrate, or        prodrug thereof, wherein L, X₁, Z, R₁, R₃, R₄, and n are defined        as for formula (I).

In another embodiment, the invention relates to compounds of formula(XI):

-   -   or a pharmaceutically acceptable salt, solvate, clathrate, or        prodrug thereof, wherein:    -   Z, R₃, R₁₈ and n are defined as for formula (I); and    -   Y is an optionally substituted 5- or 6-membered heteroaryl.

In another embodiment, the invention relates to compounds of formula(XII):

-   -   or a pharmaceutically acceptable salt, solvate, clathrate, or        prodrug thereof, wherein:    -   Z, R₃, R₁₈ and n are defined as for formula (I); and    -   R₁₉ is H, a halo, an optionally substituted alkyl, an optionally        substituted alkoxy, or an optionally substituted alkyl sulfanyl.

A compound of the invention or a pharmaceutically acceptable salt,solvate, clathrate, or prodrug thereof is particularly useful inhibitingimmune cell (e.g., T-cells, B-cells and/or mast cells) activation (e.g.,activation in response to an antigen). In particular, a compound of theinvention or a pharmaceutically acceptable salt, solvate, clathrate, orprodrug thereof can inhibit the production of certain cytokines thatregulate immune cell activation. For example, a compound of theinvention or a pharmaceutically acceptable salt, solvate, clathrate, orprodrug thereof can inhibit the production of IL-2, IL-4, IL-5, IL-13,GM-CSF, TNF-α, INF-γ or combinations thereof. Moreover, a compound ofthe invention or a pharmaceutically acceptable salt, solvate, clathrate,or prodrug thereof can modulate the activity of one or more ion channelinvolved in activation of immune cells, such as CRAC ion channels.

A compound of the invention or a pharmaceutically acceptable salt,solvate, clathrate, or prodrug thereof is particularly useful forimmunosuppression or for treating or preventing inflammatory conditions,allergic disorders, and immune disorders.

The invention also encompasses pharmaceutical compositions comprising acompound of the invention or a pharmaceutically acceptable salt,solvate, clathrate, or prodrug thereof; and a pharmaceuticallyacceptable carrier or vehicle. These compositions may further compriseadditional agents. These compositions are useful for immunosuppressionand treating or preventing inflammatory conditions, allergic disordersand immune disorders.

The invention further encompasses methods for treating or preventinginflammatory conditions, allergic disorders, and immune disorders,comprising administering to a subject in need thereof an effectiveamount of a compound of the invention or a pharmaceutically acceptablesalt, solvate, clathrate, or prodrug thereof, or a pharmaceuticalcomposition comprising a compound of the invention or a pharmaceuticallyacceptable salt, solvate, clathrate, or prodrug thereof. These methodsmay also comprise administering to the subject an additional agentseparately or in a combination composition with the compound of theinvention or a pharmaceutically acceptable salt, solvate, clathrate, orprodrug thereof.

The invention further encompasses methods for suppressing the immunesystem of a subject in need thereof, comprising administering to asubject in need thereof an effective amount of a compound of theinvention or a pharmaceutically acceptable salt, solvate, clathrate, orprodrug thereof, or a pharmaceutical composition comprising a compoundof the invention or a pharmaceutically acceptable salt, solvate,clathrate, or prodrug thereof. In one embodiment, the subject in need ofimmune system suppression is an organ transplant recipient, such as arecipient of a heart, kidney, lung, liver, skin graft, islet ofLangerhans, and the like. These methods may also comprise administeringto the subject an additional agent separately or in a combinationcomposition with the compound of the invention or a pharmaceuticallyacceptable salt, solvate, clathrate, or prodrug thereof.

The invention further encompasses methods for inhibiting immune cellactivation, including inhibiting proliferation of T cells and/or Bcells, in vivo or in vitro comprising administering to the cell aneffective amount of a compound of the invention or a pharmaceuticallyacceptable salt, solvate, clathrate, or prodrug thereof or apharmaceutical composition comprising a compound of the invention or apharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

The invention further encompasses methods for inhibiting mast celldegranulation, in vivo or in vitro comprising administering to the cellan effective amount of a compound of the invention or a pharmaceuticallyacceptable salt, solvate, clathrate, or prodrug thereof or apharmaceutical composition comprising a compound of the invention or apharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

The invention further encompasses methods for inhibiting cytokineproduction in a cell, (e.g., IL-2, IL-4, IL-5, IL-13, GM-CSF, TNF-α,and/or INF-γ production) in vivo or in vitro comprising administering toa cell an effective amount of a compound of the invention or apharmaceutically acceptable salt, solvate, clathrate, or prodrug thereofor a pharmaceutical composition comprising a compound of the inventionor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

The invention further encompasses methods for modulating ion channelactivity (e.g., CRAC) in vivo or in vitro comprising administering aneffective amount of a compound of the invention or a pharmaceuticallyacceptable salt, solvate, clathrate, or prodrug thereof or apharmaceutical composition comprising a compound of the invention or apharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

All of the methods of this invention may be practice with a compound ofthe invention alone, or in combination with other agents, such as otherimmunosuppressive agents, anti-inflammatory agents, agents for thetreatment of allergic disorders or agents for the treatment of immunedisorders.

DESCRIPTION OF THE DRAWING

FIG. 1 is a graph showing the inhibition of chemotaxis in human and minipig T cells after exposure to Compound 1.

DETAILED DESCRIPTION OF THE INVENTION Definitions

Unless otherwise specified, the below terms used herein are defined asfollows:

As used herein, the term an “aromatic ring” or “aryl” means a monocyclicor polycyclic-aromatic ring or ring radical comprising carbon andhydrogen atoms. Examples of suitable aryl groups include, but are notlimited to, phenyl, tolyl, anthacenyl, fluorenyl, indenyl, azulenyl, andnaphthyl, as well as benzo-fused carbocyclic moieties such as5,6,7,8-tetrahydronaphthyl. An aryl group can be unsubstituted orsubstituted with one or more substituents (including without limitationalkyl (preferably, lower alkyl or alkyl substituted with one or morehalo), hydroxy, alkoxy (preferably, lower alkoxy), alkylthio, cyano,halo, amino, and nitro. In certain embodiments, the aryl group is amonocyclic ring, wherein the ring comprises 6 carbon atoms.

As used herein, the term “alkyl” means a saturated straight chain orbranched non-cyclic hydrocarbon typically having from 1 to 10 carbonatoms. Representative saturated straight chain alkyls include methyl,ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyland n-decyl; while saturated branched alkyls include isopropyl,sec-butyl, isobutyl, tert-butyl, isopentyl, 2-methylbutyl,3-methylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl,2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl,2,3-dimethylbutyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl,2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl,2,2-dimethylpentyl, 2,2-dimethylhexyl, 3,3-dimethylpentyl,3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylpentyl, 3-ethylpentyl,2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl,2-methyl-3-ethylpentyl, 2-methyl-4-ethylpentyl, 2-methyl-2-ethylhexyl,2-methyl-3-ethylhexyl, 2-methyl-4-ethylhexyl, 2,2-diethylpentyl,3,3-diethylhexyl, 2,2-diethylhexyl, 3,3-diethylhexyl and the like. Alkylgroups included in compounds of this invention may be optionallysubstituted with one or more substituents, such as amino, alkylamino,alkoxy, alkylthio, oxo, halo, acyl, nitro, hydroxyl, cyano, aryl,alkylaryl, aryloxy, arylthio, arylamino, carbocyclyl, carbocyclyloxy,carbocyclylthio, carbocyclylamino, heterocyclyl, heterocyclyloxy,heterocyclylamino, heterocyclylthio, and the like. In addition, anycarbon in the alkyl segment may be substituted with oxygen (═O), sulfur(═S), or nitrogen (═NR²³, wherein R²³ is —H, an alkyl, acetyl, oraralkyl). Lower alkyls are typically preferred for the compounds of thisinvention.

The term alkylene refers to an alkyl group that has two points ofattachment to two moieties (e.g., {—CH₂—}, —{CH₂CH₂—},

etc., wherein the brackets indicate the points of attachment). Alkylenegroups may be substituted or unsubstituted.

An aralkyl group refers to an aryl group that is attached to anothermoiety via an alkylene linker. Aralkyl groups can be substituted orunsubstituted.

The term “alkoxy,” as used herein, refers to an alkyl group which islinked to another moiety though an oxygen atom. Alkoxy groups can besubstituted or unsubstituted.

The term “alkoxyalkoxy,” as used herein, refers to an alkoxy group inwhich the alkyl portion is substituted with another alkoxy group.

The term “alkyl sulfanyl,” as used herein, refers to an alkyl groupwhich is linked to another moiety though a divalent sulfur atom. Alkylsulfanyl groups can be substituted or unsubstituted.

The term “alkylamino,” as used herein, refers to an amino group in whichone hydrogen atom attached to the nitrogen has been replaced by an alkylgroup. The term “dialkylamino,” as used herein, refers to an amino groupin which two hydrogen atoms attached to the nitrogen have been replacedby alkyl groups, in which the alkyl groups can be the same or different.Alkylamino groups and dialkylamino groups can be substituted orunsubstituted.

As used herein, the term “alkenyl” means a straight chain or branched,hydrocarbon radical typically having from 2 to 10 carbon atoms andhaving at least one carbon-carbon double bond. Representative straightchain and branched alkenyls include vinyl, allyl, 1-butenyl, 2-butenyl,isobutylenyl, 1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl,1-methyl-2-butenyl, 2,3-dimethyl-2-butenyl, 1-hexenyl, 2-hexenyl,3-hexenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 1-octenyl, 2-octenyl,3-octenyl, 1-nonenyl, 2-nonenyl, 3-nonenyl, 1-decenyl, 2-decenyl,3-decenyl and the like. Alkenyl groups can be substituted orunsubstituted.

As used herein, the term “alkynyl” means a straight chain or branched,hydrocarbon radical typically having from 2 to 10 carbon atoms andhaving at lease one carbon-carbon triple bond. Representative straightchain and branched alkynyls include acetylenyl, propynyl, 1-butynyl,2-butynyl, 1-pentynyl, 2-pentynyl, 3-methyl-1-butynyl, 4-pentynyl,-1-hexynyl, 2-hexynyl, 5-hexynyl, 1-heptynyl, 2-heptynyl, 6-heptynyl,1-octynyl, 2-octynyl, 7-octynyl, 1-nonynyl, 2-nonynyl, 8-nonynyl,1-decynyl, 2-decynyl, 9-decynyl and the like. Alkynyl groups can besubstituted or unsubstituted.

As used herein, the term “cycloalkyl” means a saturated, mono- orpolycyclic alkyl radical typically having from 3 to 10 carbon atoms.Representative cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, adamantly,decahydronaphthyl, octahydropentalene, bicycle[1.1.1]pentanyl, and thelike. Cycloalkyl groups can be substituted or unsubstituted.

As used herein, the term “cycloalkenyl” means a cyclic non-aromaticalkenyl radical having at least one carbon-carbon double bond in thecyclic system and typically having from 5 to 10 carbon atoms.Representative cycloalkenyls include cyclopentenyl, cyclopentadienyl,cyclohexenyl, cyclohexadienyl, cycloheptenyl, cycloheptadienyl,cycloheptatrienyl, cyclooctenyl, cyclooctadienyl, cyclooctatrienyl,cyclooctatetraenyl, cyclononenyl, cyclononadienyl, cyclodecenyl,cyclodecadienyl and the like. Cycloalkenyl groups can be substituted orunsubstituted.

As used herein, the term “heterocycle” or “heterocyclyl” means amonocyclic or polycyclic heterocyclic ring (typically having 3- to14-members) which is either a saturated ring or a unsaturatednon-aromatic ring. A 3-membered heterocycle can contain up to 3heteroatoms, and a 4- to 14-membered heterocycle can contain from 1 toabout 8 heteroatoms. Each heteroatom is independently selected fromnitrogen, which can be quaternized; oxygen; and sulfur, includingsulfoxide and sulfone. The heterocycle may be attached via anyheteroatom or carbon atom. Representative heterocycles includemorpholinyl, thiomorpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl,piperazinyl, hydantoinyl, valerolactamyl, oxiranyl, oxetanyl,tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyrindinyl,tetrahydropyrimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, andthe like. A heteroatom may be substituted with a protecting group knownto those of ordinary skill in the art, for example, the hydrogen on anitrogen may be substituted with a tert-butoxycarbonyl group.Furthermore, the heterocyclyl may be optionally substituted with one ormore substituents (including without limitation a halogen atom, an alkylradical, or aryl radical). Only stable isomers of such substitutedheterocyclic groups are contemplated in this definition. Heterocyclylgroups can be substituted or unsubstituted.

As used herein, the term “heteroaromatic” or “heteroaryl” means amonocyclic or polycyclic heteroaromatic ring (or radical thereof)comprising carbon atom ring members and one or more heteroatom ringmembers (such as, for example, oxygen, sulfur or nitrogen). Typically,the heteroaromatic ring has from 5 to about 14 ring members in which atleast 1 ring member is a heteroatom selected from oxygen, sulfur andnitrogen. In another embodiment, the heteroaromatic ring is a 5 or 6membered ring and may contain from 1 to about 4 heteroatoms. In anotherembodiment, the heteroaromatic ring system has a 7 to 14 ring membersand may contain from 1 to about 7 heteroatoms. Representativeheteroaryls include pyridyl, furyl, thienyl, pyrrolyl, oxazolyl,imidazolyl, indolizinyl, thiazolyl, isoxazolyl, pyrazolyl, isothiazolyl,pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, triazolyl, pyridinyl,thiadiazolyl, pyrazinyl, quinolyl, isoquinolyl, indazolyl, benzoxazolyl,benzofuryl, benzothiazolyl, indolizinyl, imidazopyridinyl, isothiazolyl,tetrazolyl, benzimidazolyl, benzoxazolyl, benzothiazolyl,benzothiadiazolyl, benzoxadiazolyl, indolyl, tetrahydroindolyl,azaindolyl, imidazopyridyl, quinozalinyl, purinyl,pyrrolo[2,3]pyrimidyl, pyrazolo[3,4]pyrimidyl or benzo(b)thienyl and thelike. These heteroaryl groups may be optionally substituted with one ormore substituents

A heteroaralkyl group refers to a heteroaryl group that is attached toanother moiety via an alkylene linker. Heteroaralkyl groups can besubstituted or unsubstituted.

As used herein, the term “halogen” or “halo” means —F, —Cl, —Br or —I.

As used herein, the term “haloalkyl” means an alkyl group in which oneor more —H is replaced with a halo group. Examples of haloalkyl groupsinclude —CF₃, —CHF₂, —CCl₃, —CH₂CH₂Br, —CH₂CH(CH₂CH₂Br)CH₃, —CHICH₃, andthe like.

As used herein, the term “haloalkoxy” means an alkoxy group in which oneor more —H is replaced with a halo group. Examples of haloalkoxy groupsinclude —OCF₃ and —OCHF₂.

The terms “bioisostere” and “bioisosteric replacement” have the samemeanings as those generally recognized in the art. Bioisosteres areatoms, ions, or molecules in which the peripheral layers of electronscan be considered substantially identical. The term bioisostere isusually used to mean a portion of an overall molecule, as opposed to theentire molecule itself. Bioisosteric replacement involves using onebioisostere to replace another with the expectation of maintaining orslightly modifying the biological activity of the first bioisostere. Thebioisosteres in this case are thus atoms or groups of atoms havingsimilar size, shape and electron density. Preferred bioisosteres ofesters, amides or carboxylic acids are compounds containing two sitesfor hydrogen bond acceptance. In one embodiment, the ester, amide orcarboxylic acid bioisostere is a 5-membered monocyclic heteroaryl ring,such as an optionally substituted 1H-imidazolyl, an optionallysubstituted oxazolyl, an optionally substituted thiazolyl,1H-tetrazolyl, [1,2,4]triazolyl, or an optionally substituted[1,2,4]oxadiazolyl.

As used herein, the terms “subject”, “patient” and “animal”, are usedinterchangeably and include, but are not limited to, a cow, monkey,horse, sheep, pig, chicken, turkey, quail, cat, dog, mouse, rat, rabbit,guinea pig and human. The preferred subject, patient or animal is ahuman.

As used herein, the term “lower” refers to a group having up to fourcarbon atoms. For example, a “lower alkyl” refers to an alkyl radicalhaving from 1 to 4 carbon atoms, and a “lower alkenyl” or “loweralkynyl” refers to an alkenyl or alkynyl radical having from 2 to 4carbon atoms, respectively. A lower alkoxy or a lower alkyl sulfanylrefers to an alkoxy or a alkyl sulfanyl having from 1 to 4 carbon atoms.Lower substituents are typically preferred.

Where a particular substituent, such as an alkyl substituent, occursmultiple times in a given structure or moeity, the identity of thesubstitutent is independent in each case and may be the same as ordifferent from other occurrences of that substituent in the structure ormoiety. Furthermore, individual substituents in the specific embodimentsand exemplary compounds of this invention are preferred in combinationwith other such substituents in the compounds of this invention, even ifsuch individual substituents are not expressly noted as being preferredor not expressly shown in combination with other substituents.

The compounds of the invention are defined herein by their chemicalstructures and/or chemical names. Where a compound is referred to byboth a chemical structure and a chemical name, and the chemicalstructure and chemical name conflict, the chemical structure isdeterminative of the compound's identity.

Suitable substituents for an alkyl, alkoxy, alkyl sulfanyl, alkylamino,dialkylamino, alkylene, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,heterocyclyl, aryl, aralkyl, heteroaryl, and heteroarylalkyl groupsinclude any substituent which will form a stable compound of theinvention. Examples of substituents for an alkyl, alkoxy, alkylsulfanyl,alkylamino, dialkylamino, alkylene, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, heterocyclyl, aryl, aralkyl, heteroaryl, andheteroarylalkyl include an alkyl, alkoxy, alkyl sulfanyl, alkylamino,dialkylamino, an alkenyl, an alkynyl, an cycloalkyl, an cycloalkenyl, anheterocyclyl, an aryl, an heteroaryl, an aralkyl, an heteraralkyl, ahaloalkyl, —C(O)NR₁₃R₁₄, —NR₁₅C(O)R₁₆, halo, —OR₁₅, cyano, nitro,haloalkoxy, —C(O)R₁₅, —SR₁₅, —C(O)OR₁₅, —OC(O)OR₁₅, —OC(O)R₁₅,—NR₁₅C(O)NR₁₃R₁₄, —NR₁₅C(NR₁₆)NR₁₃R₁₄, —OC(O)NR₁₃R₁₄, —NR₁₅C(O)OR₁₆,—S(O)_(p)R₁₅, —NR₁₆S(O)_(p)R₁₅, or —S(O)_(p)NR₁₃R₁₄, wherein R₁₃ andR₁₄, for each occurrence are, independently, H, an optionallysubstituted alkyl, an optionally substituted alkenyl, an optionallysubstituted alkynyl, an optionally substituted cycloalkyl, an optionallysubstituted cycloalkenyl, an optionally substituted heterocyclyl, anoptionally substituted aryl, an optionally substituted heteroaryl, anoptionally substituted aralkyl, or an optionally substitutedheteraralkyl; or R₁₃ and R₁₄ taken together with the nitrogen to whichthey are attached is optionally substituted heterocyclyl or optionallysubstituted heteroaryl; and R₁₅ and R₁₆ for each occurrence are,independently, H, an optionally substituted alkyl, an optionallysubstituted alkenyl, an optionally substituted alkynyl, an optionallysubstituted cycloalkyl, an optionally substituted cycloalkenyl, anoptionally substituted heterocyclyl, an optionally substituted aryl, anoptionally substituted heteroaryl, an optionally substituted aralkyl, oran optionally substituted heteraralkyl.

In addition, alkyl, cycloalkyl, alkylene, a heterocyclyl, and anysaturated portion of a alkenyl, cycloalkenyl, alkynyl, aralkyl, andheteroaralkyl groups, may also be substituted with ═O, ═S, ═N—R₁₅.

When a heterocyclyl, heteroaryl, or heteroaralkyl group contains anitrogen atom, it may be substituted or unsubstituted. When a nitrogenatom in the aromatic ring of a heteroaryl group has a substituent thenitrogen may be a quaternary nitrogen.

Choices and combinations of substituents and variables envisioned bythis invention are only those that result in the formation of stablecompounds. The term “stable”, as used herein, refers to compounds whichpossess stability sufficient to allow manufacture and which maintainsthe integrity of the compound for a sufficient period of time to beuseful for the purposes detailed herein (e.g., therapeutic orprophylactic administration to a subject). Typically, such compounds arestable at a temperature of 40° C. or less, in the absence of excessivemoisture, for at least one week. Such choices and combinations will beapparent to those of ordinary skill in the art and may be determinedwithout undue experimentation.

Unless indicated otherwise, the compounds of the invention containingreactive functional groups (such as, without limitation, carboxy,hydroxy, and amino moieties) also include protected derivatives thereof.“Protected derivatives” are those compounds in which a reactive site orsites are blocked with one ore more protecting groups. Suitableprotecting groups for carboxy moieties include benzyl, tert-butyl, andthe like. Suitable protecting groups for amino and amido groups includeacetyl, tert-butoxycarbonyl, benzyloxycarbonyl, and the like. Suitableprotecting groups for hydroxy include benzyl and the like. Othersuitable protecting groups are well known to those of ordinary skill inthe art and include those found in T. W. Greene, Protecting Groups inOrganic Synthesis, John Wiley & Sons, Inc. 1981, the entire teachings ofwhich are incorporated herein by reference.

As used herein, the term “compound(s) of this invention” and similarterms refers to a compound of any one of formulas (I) through (XII), orTable 1, or a pharmaceutically acceptable salt, solvate, clathrate, orprodrug thereof and also include protected derivatives thereof.

As used herein and unless otherwise indicated, the term “prodrug” meansa derivative of a compound that can hydrolyze, oxidize, or otherwisereact under biological conditions (in vitro or in vivo) to provide acompound of this invention. Prodrugs may only become active upon suchreaction under biological conditions, but they may have activity intheir unreacted forms. Examples of prodrugs contemplated in thisinvention include, but are not limited to, analogs or derivatives ofcompounds of any one of formulas (I) through (XII), or Table 1 thatcomprise biohydrolyzable moieties such as biohydrolyzable amides,biohydrolyzable esters, biohydrolyzable carbamates, biohydrolyzablecarbonates, biohydrolyzable ureides, and biohydrolyzable phosphateanalogues. Other examples of prodrugs include derivatives of compoundsof any one of formulas (I) through (XII), or of Table 1 that comprise—NO, —NO₂, —ONO, or —ONO₂ moieties. Prodrugs can typically be preparedusing well-known methods, such as those described by 1 BURGER'SMEDICINAL CHEMISTRY AND DRUG DISCOVERY (1995) 172-178, 949-982 (ManfredE. Wolff ed., 5^(th) ed), the entire teachings of which are incorporatedherein by reference.

As used herein and unless otherwise indicated, the terms“biohydrolyzable amide”, “biohydrolyzable ester”, “biohydrolyzablecarbamate”, “biohydrolyzable carbonate”, “biohydrolyzable ureide” and“biohydrolyzable phosphate analogue” mean an amide, ester, carbamate,carbonate, ureide, or phosphate analogue, respectively, that either: 1)does not destroy the biological activity of the compound and confersupon that compound advantageous properties in vivo, such as uptake,duration of action, or onset of action; or 2) is itself biologicallyinactive but is converted in vivo to a biologically active compound.Examples of biohydrolyzable amides include, but are not limited to,lower alkyl amides, α-amino acid amides, alkoxyacyl amides, andalkylaminoalkylcarbonyl amides. Examples of biohydrolyzable estersinclude, but are not limited to, lower alkyl esters, alkoxyacyloxyesters, alkyl acylamino alkyl esters, and choline esters. Examples ofbiohydrolyzable carbamates include, but are not limited to, loweralkylamines, substituted ethylenediamines, aminoacids,hydroxyalkylamines, heterocyclic and heteroaromatic amines, andpolyether amines.

As used herein, the term “pharmaceutically acceptable salt,” is a saltformed from an acid and a basic group of one of the compounds of any oneof formulas (I) through (XII) or of Table 1. Illustrative salts include,but are not limited, to sulfate, citrate, acetate, oxalate, chloride,bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate,isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate,tannate, pantothenate, bitartrate, ascorbate, succinate, maleate,gentisinate, fumarate, gluconate, glucaronate, saccharate, formate,benzoate, glutamate, methanesulfonate, ethanesulfonate,benzenesulfonate, p-toluenesulfonate, and pamoate (i.e.,1,1′-methylene-bis-(2-hydroxy-3-naphthoate)) salts. The term“pharmaceutically acceptable salt” also refers to a salt prepared from acompound of any one of formulas (I) through (XII) or Table 1 having anacidic functional group, such as a carboxylic acid functional group, anda pharmaceutically acceptable inorganic or organic base. Suitable basesinclude, but are not limited to, hydroxides of alkali metals such assodium, potassium, and lithium; hydroxides of alkaline earth metal suchas calcium and magnesium; hydroxides of other metals, such as aluminumand zinc; ammonia, and organic amines, such as unsubstituted orhydroxy-substituted mono-, di-, or trialkylamines; dicyclohexylamine;tributyl amine; pyridine; N-methyl,N-ethylamine; diethylamine;triethylamine; mono-, bis-, or tris-(2-hydroxy-lower alkyl amines), suchas mono-, bis-, or tris-(2-hydroxyethyl)-amine,2-hydroxy-tert-butylamine, or tris-(hydroxymethyl)methylamine,N,N,-di-lower alkyl-N-(hydroxy lower alkyl)-amines, such asN,N-dimethyl-N-(2-hydroxyethyl)amine, or tri-(2-hydroxyethyl)amine;N-methyl-D-glucamine; and amino acids such as arginine, lysine, and thelike. The term “pharmaceutically acceptable salt” also refers to a saltprepared from a compound of any one of formulas (I) through (XII) orTable 1 having a basic functional group, such as an amino functionalgroup, and a pharmaceutically acceptable inorganic or organic acid.Suitable acids include, but are not limited to, hydrogen sulfate, citricacid, acetic acid, oxalic acid, hydrochloric acid, hydrogen bromide,hydrogen iodide, nitric acid, phosphoric acid, isonicotinic acid, lacticacid, salicylic acid, tartaric acid, ascorbic acid, succinic acid,maleic acid, besylic acid, fumaric acid, gluconic acid, glucaronic acid,saccharic acid, formic acid, benzoic acid, glutamic acid,methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, andp-toluenesulfonic acid.

As used herein, the term “pharmaceutically acceptable solvate,” is asolvate formed from the association of one or more solvent molecules toone or more molecules of a compound of any one of formulas (I) through(XII) or Table 1. The term solvate includes hydrates (e.g.,hemi-hydrate, mono-hydrate, dihydrate, trihydrate, tetrahydrate, and thelike).

As used herein, the term “clathrate” means a compound of the presentinvention or a salt thereof in the form of a crystal lattice thatcontains spaces (e.g., channels) that have a guest molecule (e.g., asolvent or water) trapped within.

As used herein, the term “asthma” means a pulmonary disease, disorder orcondition characterized by reversible airway obstruction, airwayinflammation, and increased airway responsiveness to a variety ofstimuli.

“Immunosuppression” refers to impairment of any component of the immunesystem resulting in decreased immune function. This impairment may bemeasured by any conventional means including whole blood assays oflymphocyte function, detection of lymphocyte proliferation andassessment of the expression of T cell surface antigens. The antisheepred blood cell (SRBC) primary (IgM) antibody response assay (usuallyreferred to as the plaque assay) is one specific method. This and othermethods are described in Luster, M. I., Portier, C., Pait, D. G., White,K. L., Jr., Gennings, C., Munson, A. E., and Rosenthal, G. J. (1992).“Risk Assessment in Immunotoxicology I: Sensitivity and Predictabilityof Immune Tests.” Fundam. Appl. Toxicol., 18, 200-210. Measuring theimmune response to a T-cell dependent immunogen is another particularlyuseful assay (Dean, J. H., House, R. V., and Luster, M. I. (2001).“Immunotoxicology: Effects of, and Responses to, Drugs and Chemicals.”In Principles and Methods of Toxicology: Fourth Edition (A. W. Hayes,Ed.), pp. 1415-1450, Taylor & Francis, Philadelphia, Pa.).

A subject in need of immune system suppression, as used herein, is asubject that is about to undergo or has had an organ transplant or asubject that has an inflammatory disorder, an immune disorder or anallergic disorder or is at risk of the reoccurrence of inflammatorydisorder, an immune disorder or an allergic disorder. In one embodiment,a subject in need of immune system suppression is at risk of aquiring ordeveloping an inflammatory disorder, an immune disorder or an allergicdisorder based on, for example, the subject's medical history or geneticbackground. The risk of the reoccurrence of or of aquiring or developingan inflammatory disorder, an immune disorder or an allergic disorder iswithin the judgement of a physician skilled in the art.

The compounds of this invention can be used to treat subjects withimmune disorders. As used herein, the term “immune disorder” and liketerms means a disease, disorder or condition caused by the immune systemof an animal, including autoimmune disorders. Immune disorders includethose diseases, disorders or conditions that have an immune componentand those that are substantially or entirely immune system-mediated.Autoimmune disorders are those wherein the animal's own immune systemmistakenly attacks itself, thereby targeting the cells, tissues, and/ororgans of the animal's own body. For example, the autoimmune reaction isdirected against the nervous system in multiple sclerosis and the gut inCrohn's disease. In other autoimmune disorders such as systemic lupuserythematosus (lupus), affected tissues and organs may vary amongindividuals with the same disease. One person with lupus may haveaffected skin and joints whereas another may have affected skin, kidney,and lungs. Ultimately, damage to certain tissues by the immune systemmay be permanent, as with destruction of insulin-producing cells of thepancreas in Type 1 diabetes mellitus. Specific autoimmune disorders thatmay be ameliorated using the compounds and methods of this inventioninclude without limitation, autoimmune disorders of the nervous system(e.g., multiple sclerosis, myasthenia gravis, autoimmune neuropathiessuch as Guillain-Barré, and autoimmune uveitis), autoimmune disorders ofthe blood (e.g., autoimmune hemolytic anemia, pernicious anemia, andautoimmune thrombocytopenia), autoimmune disorders of the blood vessels(e.g., temporal arteritis, anti-phospholipid syndrome, vasculitides suchas Wegener's granulomatosis, and Behcet's disease), autoimmune disordersof the skin (e.g., psoriasis, dermatitis herpetiformis, pemphigusvulgaris, and vitiligo), autoimmune disorders of the gastrointestinalsystem (e.g., Crohn's disease, ulcerative colitis, primary biliarycirrhosis, and autoimmune hepatitis), autoimmune disorders of theendocrine glands (e.g., Type 1 or immune-mediated diabetes mellitus,Grave's disease. Hashimoto's thyroiditis, autoimmune oophoritis andorchitis, and autoimmune disorder of the adrenal gland); and autoimmunedisorders of multiple organs (including connective tissue andmusculoskeletal system diseases) (e.g., rheumatoid arthritis, systemiclupus erythematosus, scleroderma, polymyositis, dermatomyositis,spondyloarthropathies such as ankylosing spondylitis, and Sjogren'ssyndrome). In addition, other immune system mediated diseases, such asgraft-versus-host disease and allergic disorders, are also included inthe definition of immune disorders herein. Because a number of immunedisorders are caused by inflammation, there is some overlap betweendisorders that are considered immune disorders and inflammatorydisorders. For the purpose of this invention, in the case of such anoverlapping disorder, it may be considered either an immune disorder oran inflammatory disorder. “Treatment of an immune disorder” hereinrefers to administering a compound or a composition of the invention toa subject, who has an immune disorder, a symptom of such a disease or apredisposition towards such a disease, with the purpose to cure,relieve, alter, affect, or prevent the autoimmune disorder, the symptomof it, or the predisposition towards it.

As used herein, the term “allergic disorder” means a disease, conditionor disorder associated with an allergic response against normallyinnocuous substances. These substances may be found in the environment(such as indoor air pollutants and aeroallergens) or they may benon-environmental (such as those causing dermatological or foodallergies). Allergens can enter the body through a number of routes,including by inhalation, ingestion, contact with the skin or injection(including by insect sting). Many allergic disorders are linked toatopy, a predisposition to generate the allergic antibody IgE. BecauseIgE is able to sensitize mast cells anywhere in the body, atopicindividuals often express disease in more than one organ. For thepurpose of this invention, allergic disorders include anyhypersensitivity that occurs upon re-exposure to the sensitizingallergen, which in turn causes the release of inflammatory mediators.Allergic disorders include without limitation, allergic rhinitis (e.g.,hay fever), sinusitis, rhinosinusitis, chronic or recurrent otitismedia, drug reactions, insect sting reactions, latex reactions,conjunctivitis, urticaria, anaphylaxis and anaphylactoid reactions,atopic dermatitis, asthma and food allergies.

The compounds of this invention can be used to prevent or to treatsubjects with inflammatory disorders. As used herein, an “inflammatorydisorder” means a disease, disorder or condition characterized byinflammation of body tissue or having an inflammatory component. Theseinclude local inflammatory responses and systemic inflammation. Examplesof such inflammatory disorders include: transplant rejection, includingskin graft rejection; chronic inflammatory disorders of the joints,including arthritis, rheumatoid arthritis, osteoarthritis and bonediseases associated with increased bone resorption; inflammatory boweldiseases such as ileitis, ulcerative colitis, Barrett's syndrome, andCrohn's disease; inflammatory lung disorders such as asthma, adultrespiratory distress syndrome, and chronic obstructive airway disease;inflammatory disorders of the eye including corneal dystrophy, trachoma,onchocerciasis, uveitis, sympathetic ophthalmitis and endophthalmitis;chronic inflammatory disorders of the gums, including gingivitis andperiodontitis; tuberculosis; leprosy; inflammatory diseases of thekidney including uremic complications, glomerulonephritis and nephrosis;inflammatory disorders of the skin including sclerodermatitis, psoriasisand eczema; inflammatory diseases of the central nervous system,including chronic demyelinating diseases of the nervous system, multiplesclerosis, AIDS-related neurodegeneration and Alzheimer's disease,infectious meningitis, encephalomyelitis, Parkinson's disease,Huntington's disease, amyotrophic lateral sclerosis and viral orautoimmune encephalitis; autoimmune disorders, immune-complexvasculitis, systemic lupus and erythematodes; systemic lupuserythematosus (SLE); and inflammatory diseases of the heart such ascardiomyopathy, ischemic heart disease hypercholesterolemia,atherosclerosis); as well as various other diseases with significantinflammatory components, including preeclampsia; chronic liver failure,brain and spinal cord trauma, cancer). There may also be a systemicinflammation of the body, exemplified by gram-positive or gram negativeshock, hemorrhagic or anaphylactic shock, or shock induced by cancerchemotherapy in response to pro-inflammatory cytokines, e.g., shockassociated with pro-inflammatory cytokines. Such shock can be induced,e.g., by a chemotherapeutic agent used in cancer chemotherapy.“Treatment of an inflammatory disorder” herein refers to administering acompound or a composition of the invention to a subject, who has aninflammatory disorder, a symptom of such a disorder or a predispositiontowards such a disorder, with the purpose to cure, relieve, alter,affect, or prevent the inflammatory disorder, the symptom of it, or thepredisposition towards it.

An “effective amount” is the quantity of compound in which a beneficialoutcome is achieved when the compound is administered to a subject oralternatively, the quantity of compound that possess a desired activityin-vivo or in-vitro. In the case of inflammatory disorders and immunedisorders, a beneficial clinical outcome includes reduction in theextent or severity of the symptoms associated with the disease ordisorder and/or an increase in the longevity and/or quality of life ofthe subject compared with the absence of the treatment. The preciseamount of compound administered to a subject will depend on the type andseverity of the disease or condition and on the characteristics of thesubject, such as general health, age, sex, body weight and tolerance todrugs. It will also depend on the degree, severity and type ofinflammatory disorder or autoimmune disorder or the degree ofimmunosuppression sought. The skilled artisan will be able to determineappropriate dosages depending on these and other factors. Effectiveamounts of the disclosed compounds typically range between about 1mg/mm² per day and about 10 grams/mm² per day, and preferably between 10mg/mm² per day and about 1 gram/mm².

The compounds of the invention may contain one or more chiral centersand/or double bonds and, therefore, exist as stereoisomers, such asdouble-bond isomers (i.e., geometric isomers), enantiomers, ordiastereomers. According to this invention, the chemical structuresdepicted herein, including the compounds of this invention, encompassall of the corresponding compounds' enantiomers and stereoisomers, thatis, both the stereomerically pure form (e.g., geometrically pure,enantiomerically pure, or diastereomerically pure) and enantiomeric,diastereomeric, and geometric isomeric mixtures. In some cases, oneenantiomer, diastereomer, or geometric isomer will possess superioractivity or an improved toxicity or kinetic profile compared to others.In those cases, such enantiomers, diastereomers, and geometric isomersof a compound of this invention are preferred.

The term “inhibit production of IL-2” and like terms means inhibitingIL-2 synthesis (e.g. by inhibiting transcription (mRNA expression), ortranslation (protein expression)) and/or inhibiting IL-2 secretion in acell that has the ability to produce and/or secrete IL-2 (e.g., Tlymphocyte). Likewise, the term “inhibiting production of IL-4, IL-5,IL-13, GM-CSF, TNF-α or INF-γ means inhibiting the synthesis (e.g. byinhibiting transcription, or translation) and/or inhibiting thesecretion in a cell that has the ability to produce and/or secrete thesecytokines.

As used herein, a composition that “substantially” comprises a compoundmeans that the composition contains more than about 80% by weight, morepreferably more than about 90% by weight, even more preferably more thanabout 95% by weight, and most preferably more than about 97% by weightof the compound.

As used herein, a composition that is “substantially free” of a compoundmeans that the composition contains less than about 20% by weight, morepreferably less than about 10% by weight, even more preferably less thanabout 5% by weight, and most preferably less than about 3% by weight ofthe compound.

As used herein, a reaction that is “substantially complete” means thatthe reaction contains more than about 80% by weight of the desiredproduct, more preferably more than about 90% by weight of the desiredproduct, even more preferably more than about 95% by weight of thedesired product, and most preferably more than about 97% by weight ofthe desired product.

As used herein, a racemic mixture means about 50% of one enantiomer andabout 50% of is corresponding enantiomer relative to all chiral centersin the molecule. The invention encompasses all enantiomerically-pure,enantiomerically-enriched, diastereomerically pure, diastereomericallyenriched, and racemic mixtures of the compounds of any one of formulas(I) through (XII) or Table 1.

Enantiomeric and diastereomeric mixtures can be resolved into theircomponent enantiomers or stereoisomers by well known methods, such aschiral-phase gas chromatography, chiral-phase high performance liquidchromatography, crystallizing the compound as a chiral salt complex, orcrystallizing the compound in a chiral solvent. Enantiomers anddiastereomers can also be obtained from diastereomerically- orenantiomerically-pure intermediates, reagents, and catalysts by wellknown asymmetric synthetic methods.

When administered to a patient, e.g., to a non-human animal forveterinary use or for improvement of livestock, or to a human forclinical use, the compounds of the invention are typically administeredin isolated form or as the isolated form in a pharmaceuticalcomposition. As used herein, “isolated” means that the compounds of theinvention are separated from other components of either (a) a naturalsource, such as a plant or cell, preferably bacterial culture, or (b) asynthetic organic chemical reaction mixture. Preferably, viaconventional techniques, the compounds of the invention are purified. Asused herein, “purified” means that when isolated, the isolate containsat least 95%, preferably at least 98%, of a single compound of theinvention by weight of the isolate.

Only those choices and combinations of substituents that result in astable structure are contemplated. Such choices and combinations will beapparent to those of ordinary skill in the art and may be determinedwithout undue experimentation.

The invention can be understood more fully by reference to the followingdetailed description and illustrative examples, which are intended toexemplify non-limiting embodiments of the invention.

SPECIFIC EMBODIMENTS

The invention relates to compounds and pharmaceutical compositions thatare particularly useful for immunosuppression or to treat or preventinflammatory conditions, immune disorders, and allergic disorders.

One embodiment of the invention relates to compounds of formula (I):

-   -   or a pharmaceutically acceptable salt, solvate, clathrate, or        prodrug thereof, wherein:    -   L is a linker selected from the group consisting of a covalent        bond, —NRCH₂—, —CH₂NR—, —C(O)—, —NR—C(O)—, —C(O)—NR—, —OC(O)—,        —C(O)O—, —C(S)—, —NR—C(S)—, —C(S)—NR—;    -   X₁ and X₃ are each, independently, CH or N;    -   X₂ is CH, CR₁₀ or N;    -   each Z is independently selected from the group consisting of a        lower alkyl, a lower haloalkyl, a halo, a lower alkoxy, a lower        alkyl sulfanyl, cyano, nitro, or lower haloalkoxy;    -   R, for each occurrence is independently selected from —H, an        alkyl, —C(O)R₅, or —C(O)OR₅;    -   R₁ and R₂ are each, independently, a halo, a haloalkyl, a lower        alkyl, a lower alkoxy, or a haloalkoxy;    -   R₃ is an alkyl, a haloalkyl, a halo, a haloalkoxy, —OR₅, —SR₅,        or —NR₆R₇;    -   R₁₈ is a halo, cyano, nitro, —C(O)R₅, —C(O)OR₅, —C(O)SR₅,        —C(O)NR₆R₇, —C(S)R₅, —C(S)OR₅, —C(S)SR₅, —C(S)NR₆R₇, —C(NR₈)R₅,        —C(NR₈)OR₅, —C(NR₈)SR₅, —C(NR₈)NR₈R₇, —S(O)_(p)R₅, —S(O)_(p)NR₅,        —S(O)_(p)OR₅, —P(O)(OR₅)₂, —OP(O)(OR₅)₂, —P(O)(R₅)₂, a five or        six membered optionally substituted heterocycloalkyl, a five or        six membered optionally substituted heterocyclyl, or a five or        six membered optionally substituted heteroaryl;    -   R₅, for each occurrence, is independently, H, an optionally        substituted alkyl, an optionally substituted alkenyl, an        optionally substituted alkynyl, an optionally substituted        cycloalkyl, an optionally substituted cycloalkenyl, an        optionally substituted heterocyclyl, an optionally substituted        aryl, an optionally substituted heteroaryl, an optionally        substituted aralkyl, or an optionally substituted heteraralkyl;    -   R₆ and R₇, for each occurrence are, independently, H, an        optionally substituted alkyl, an optionally substituted alkenyl,        an optionally substituted alkynyl, an optionally substituted        cycloalkyl, an optionally substituted cycloalkenyl, an        optionally substituted heterocyclyl, an optionally substituted        aryl, an optionally substituted heteroaryl, an optionally        substituted aralkyl, or an optionally substituted heteraralkyl;        or R₆ and R₇ taken together with the nitrogen to which they are        attached are an optionally substituted heterocyclyl or        optionally substituted heteroaryl;    -   R₈, for each occurrence, is independently —H, a halo, an alkyl,        —OR₅, —NR₆R₇, —C(O)R₅, —C(O)OR₅, or —C(O)NR₆R₇;    -   R₁₀ is a lower alkyl, a lower alkoxy, a halo, a lower haloalkyl,        a lower haloalkoxy, a cyano, nitro, —C(O)R₅, —C(O)OR₅, —C(O)SR₅,        —C(O)NR₆R₇, —C(S)R₅, —C(S)OR₅, —C(S)SR₅, —C(S)NR₆R₇, —C(NR₈)R₅,        —C(NR₈)OR₅, —C(NR₈)SR₅, —C(NR₈)NR₆R₇, —S(O)_(p)R₅, —P(O)(OR₅)₂,        —OP(O)(OR₅)₂, or —P(O)(R₅)₂; n is zero or an integer from 1 to        4; and    -   p, for each occurrence, is independently 1 or 2.

In one embodiment of the compounds represented by formula (I), R₁₈ is anoptionally substituted pyridinyl, an optionally substituted oxazolyl, anoptionally substituted isoxazolyl, an optionally substituted pyrazolyl,an optionally substituted thiazolyl, an optionally substituted pyrrolyl,an optionally substituted morpholinyl, an optionally substitutedfuranyl, an optionally substituted thienyl, an optionally substitutedthiadiazolyl, an optionally substituted triazolyl, an optionallysubstituted oxadiazolyl, or an optionally substituted tetrazolyl.Preferably, R₁₈ is unsubstituted or substituted with one or moresubstituents selected from the group consisting of a lower alkyl, halo,a lower haloalkyl, an amino, a lower dialkyl amino, a lower alkyl amino,a lower alkoxy, and a lower alkyl sulfanyl.

In another embodiment of the compounds represented by formula (I), R₁₈is an ester, amide or carboxylic acid bioisostere. Preferably, when R₁₈is an ester, amide or carboxylic acid bioisostere, it is an optionallysubstituted oxazolyl, an optionally substituted thiazolyl, an optionallysubstituted 1H-tetrazolyl, an optionally substituted 1H-imidazolyl, anoptionally substituted [1,2,4]oxadiazolyl, or an optionally substituted4H-[1,2,4]triazolyl.

In another embodiment of the compounds represented by formula (I), R₁₈is a halo, —C(O)R₉, —S(O)_(p)R₁₁, —S(O)_(p)NR₅, —S(O)_(p)OR₅,—P(O)(OR₁₂)₂, or —P(O)(R₁₁)₂, wherein:

-   -   R₉ is a lower alkoxy, lower alkyl sulfanyl, or an alkoxyalkoxy;    -   R₁₁, for each occurrence, is independently, a lower alkyl; and    -   R₁₂, for each occurrence, is independently, H or a lower alkyl.

In another embodiment, the invention relates to compounds selected fromthe group consisting of:

-   2,6-Diffluoro-N-[2′-methyl-5′-(pyridine-3-yl)-biphenyl-4-yl]-benzamide;-   2,6-Difluoro-N-[2′-methyl-5′-(pyridine-2-yl)-biphenyl-4-yl]-benzamide;-   2,6-Difluoro-N-[2′-methyl-5′-(pyridine-4-yl)-biphenyl-4-yl]-benzamide;-   2,6-Difluoro-N-[2′-methyl-5′-(3-methyl-isoxazole-5-yl)-biphenyl-4-yl]-benzamide;-   2,6-Difluoro-N-[2′-methyl-5′-(3-methyl-1H-pyrazol-5-yl)-biphenyl-4-yl]-benzamide;-   2,6-Difluoro-N-[2′-methyl-5′-(1H-pyrrol-2-yl)-biphenyl-4-yl]-benzamide;-   2,6-Difluoro-N-[2′-methyl-5′-(5-oxo-4,5-dihydro-[1,2,4]-oxadiazol-3-yl)-biphenyl-4-yl]-benzamide;-   2,6-Difluoro-N-[2′-methyl-5′-(morpholino-4-yl)-biphenyl-4-yl]-benzamide;-   3,5-Difluoro-N-[5′-([1,3,4]thiadiazol-2-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide;-   2,6-Difluoro-N-[2′-methyl-5′-([1,3,4]thiadiazol-2-yl)-biphenyl-4-yl]-benzamide;-   2,6-Difluoro-N-[2′-methyl-5′-(5-amino-[1,3,4]thiadiazol-2-yl)-biphenyl-4-yl]-benzamide;-   2,6-Difluoro-N-{2′-methyl-5′[5-(N,N-dimethylamino)-[1,3,4]thiadiazol-2-yl]-biphenyl-4-yl}-N-methyl-benzamide;-   2,6-Difluoro-N-{2′-methyl-5′-[5-(N,N-dimethylamino)-[1,3,4]thiadiazol-2-yl]-biphenyl-4-yl}-benzamide;

and pharmaceutically acceptable salts, solvates, clathrates, andprodrugs thereof.

In another embodiment, the invention relates to compounds of formula(II):

-   -   or a pharmaceutically acceptable salt, solvate, clathrate, or        prodrug thereof, wherein:    -   L, X₁, X₂, X₃, Z, R₁, R₂, R₃ and n are defined as for formula        (I); and    -   R₄ is a halo, cyano, nitro, —C(O)R₅, —C(O)OR₅, —C(O)SR₅,        —C(O)NR₆R₇, —C(S)R₅, —C(S)OR₅, —C(S)SR₅, —C(S)NR₆R₇, —C(NR₈)R₅,        —C(NR₈)OR₅, —C(NR₈)SR₅, —C(NR₈)NR₆R₇, —S(O)_(p)R₅, —S(O)_(p)NR₅,        —S(O)_(p)OR₅, —P(O)(OR₅)₂, —OP(O)(OR₅)₂, —P(O)(R₅)₂, or an        ester, amide or carboxylic acid bioisostere.

In one embodiment of the compounds represented by formula (I) or (II), nis 0. In another embodiment, n is 1. In another embodiment, n is 2.

In another embodiment of the compounds represented by formula (I) or(II), Z, for each occurrence, is independently, a lower alkyl, a loweralkoxy, a lower haloalkoxy, a halo, cyano, or haloalkyl and n is 1 or 2.

In another embodiment of the compounds represented by formula (I) or(II), L is —NHC(O)—.

In another embodiment of the compounds represented by formula (I) or(II), L is —NHCH₂—.

In another embodiment, the invention relates to compounds of formula(III):

or a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof, wherein R₁, R₂, R₃, and R₄ are defined as above.

In another embodiment, the invention relates to compounds of formula(IV):

or a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof, wherein R₁, R₂, R₃, and R₄ are defined as above.

In another embodiment, the invention relates to compounds of formula(V):

or a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof, wherein R₁, R₂, R₃, and R₄ are defined as above.

In another embodiment, the invention relates to compounds of formula(VI):

or a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof, wherein R₁, R₂, R₃, R₄ and R₁₀ are defined as above, and

In another embodiment of the compounds represented by formula (I), (II),(III), (IV), (V), or (VI), R₁ and R₂ are each, independently, a halo.

In another embodiment of the compounds represented by formula (I), (II),(III), (IV), (V), or (VI), R₃ is a lower alkyl, a lower alkoxy, a loweralkyl sulfanyl, or a halo.

In another embodiment of the compounds represented by formula (II),(III), (IV), (V), or (VI), R₄ is a bioisostere of an ester, amide, orcarboxylic acid. For example, R₄ is a 5-membered heteroaryl, such as, anoptionally substituted oxazolyl, an optionally substituted thiazolyl, anoptionally substituted 1H-tetrazolyl, an optionally substituted1H-imidazolyl, an optionally substituted [1,2,4]oxadiazolyl, or anoptionally substituted 4H-[1,2,4]triazolyl.

In another embodiment of the compounds represented by formula (II),(III), (IV), (V), or (VI), R₄ is a halo, —C(O)R₉, —S(O)_(p)R₁₁,—S(O)_(p)NR₅, —S(O)_(p)OR₅, —P(O)(OR₁₂)₂, or —P(O)(R₁₁)₂, wherein:

-   -   R₉ is a lower alkoxy, lower alkyl sulfanyl, or an alkoxyalkoxy;    -   R₁₁, for each occurrence, is independently, a lower alkyl; and    -   R₁₂, for each occurrence, is independently, H or a lower alkyl.

In another embodiment of the compounds represented by formula (I), (II),(III), (IV), (V), or (VI), R₁ and R₂ are each a fluoro group.

In another embodiment, the invention relates to compounds selected fromthe group consisting of:

-   4′-(2,6-Difluoro-benzoylamino)-6-methyl-biphenyl-3-carboxylic acid    methyl ester;-   6-Chloro-4′-(2,6-difluoro-benzoylamino)-biphenyl-3-(carboxylic acid    2-methoxyethyl ester);-   2,6-Difluoro-N-(2′-methyl-5′-oxazol-2-yl-biphenyl-4-yl)-benzamide;-   4′-[(3,5-Difluoro-pyridine-4-cabonyl)-amino]-6-methyl-biphenyl-3-carboxylic    acid methyl ester;-   N-[4-(5-Chloro-2-methoxy-pyridin-3-yl)-phenyl]-2,6-difluoro-benzamide;-   2,6-Difluoro-N-[2′-methyl-5′-(1H-tetrazol-5-yl)-biphenyl-4-yl]-benzamide;-   3,5-Difluoro-N-(2′-methyl-5′-oxazol-2-yl-biphenyl-4-yl)-isonicotinamide;-   2,6-Difluoro-N-(2′-methoxy-5′-oxazol-2-yl-biphenyl-4-yl)-benzamide;-   2,6-Difluoro-N-[2′-methyl-5′-(oxazol-5-yl)-biphenyl-4-yl]-benzamide;-   3,5-Difluoro-N-[5′-(thiazol-2-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide;-   2,6-Difluoro-N-[2′-chloro-5′-(oxazol-2-yl)-biphenyl-4-yl]-benzamide;-   2,6-Difluoro-N-[2′-methyl-5′-(4-methyl-thiazol-2-yl)-biphenyl-4-yl]-benzamide;-   2,6-Difluoro-N-[2′-methyl-5′-(4-trifluoromethyl-thiazol-2-yl)-biphenyl-4-yl]-benzamide;-   3,5-Difluoro-N-[5′-(oxazol-2-yl)-2′-chloro-biphenyl-4-yl]-isonicotinamide;-   3,5-Difluoro-N-[5′-(oxazol-2-yl)-2′-methoxy-biphenyl-4-yl]-isonicotinamide;-   2,6-Difluoro-N-[2′-chloro-5′-(thiazol-2-yl)-biphenyl-4-yl]-benzamide;-   3,5-Difluoro-N-[5′-(thiazol-2-yl)-2′-chloro-biphenyl-4-yl]isonicotinamide;-   2,6-Difluoro-N-[2′-methyl-5′-(1H-tetrazol-5-yl)-biphenyl-4-yl]-benzamide;-   2,6-Difluoro-N-[2′-methyl-5′-(2-methyl-2H-[1,2,4]-triazol-3-yl)-biphenyl-4-yl]-benzamide;-   3,5-Difluoro-N-[2′-methyl-5′-([1,3,4]oxadiazol-2-yl)-biphenyl-4-yl]-isonicotinamide;-   2,6-Difluoro-N-[2′-methyl-5′-([1,3,4]oxadiazol-2-yl)-biphenyl-4-yl]-benzamide;-   2,6-Difluoro-N-[2′-methyl-5′-(4-methyl-5-methylsulfanyl-4H-[1,2,4]triazol-3-yl)-biphenyl-4-yl]-benzamide;-   3,5-Difluoro-N-[2′-methyl-5′-(4-methyl-5-methylsulfanyl-4H-[1,2,4]triazol-3-yl)-biphenyl-4-yl]-isonicotinamide-   2,6-Difluoro-N-[2′-methyl-5′-(oxazol-4-yl)-biphenyl-4-yl]-benzamide;-   2,6-Difluoro-N-[2′-methyl-5′-(1H-tetrazol-5-yl)-biphenyl-4-yl]-benzamide;-   2,6-Difluoro-N-[2′-methyl-5′-(1-methyl-1H-tetrazol-5-yl)-biphenyl-4-yl]-benzamide;-   2,6-Difluoro-N-[2′-methyl-5′-(2-methyl-2H-tetrazol-5-yl)-biphenyl-4-yl]-benzamide;    -   and pharmaceutically acceptable salts, solvates, clathrates, or        prodrugs thereof.

In another embodiment, the invention relates to compounds of formula(VII):

-   -   or a pharmaceutically acceptable salt, solvate, clathrate, or        prodrug thereof, wherein:    -   X₁, L, Z, R₁, R₃, R₁₈ and n are defined as for formula (I).

In one embodiment of the compounds represented by formula (VII), R₁₈ isan optionally substituted pyridinyl, an optionally substituted oxazolyl,an optionally substituted isoxazolyl, an optionally substitutedpyrazolyl, an optionally substituted thiazolyl, an optionallysubstituted pyrrolyl, an optionally substituted morpholinyl, anoptionally substituted furanyl, an optionally substituted thienyl, anoptionally substituted thiadiazolyl, an optionally substitutedtriazolyl, an optionally substituted oxadiazolyl, or an optionallysubstituted tetrazolyl. Preferably, R₁₈ is unsubstituted or substitutedwith one or more substituents selected from the group consisting of alower alkyl, halo, a lower haloalkyl, an amino, a lower dialkyl amino, alower alkyl amino, a lower alkoxy, and a lower alkyl sulfanyl.

In another embodiment of the compounds represented by formula (VII), R₁₈is an ester, amide or carboxylic acid bioisostere. Preferably, when R₁₈is an ester, amide or carboxylic acid bioisostere, it is an optionallysubstituted oxazolyl, an optionally substituted thiazolyl, an optionallysubstituted 1H-tetrazolyl, an optionally substituted 1H-imidazolyl, anoptionally substituted [1,2,4]oxadiazolyl, or an optionally substituted4H-[1,2,4]triazolyl.

In another embodiment of the compounds represented by formula (VII), R₁₈is a halo, —C(O)R₉, —S(O)_(p)R₁₁, —S(O)_(p)NR₅, —S(O)_(p)OR₅,—P(O)(OR₁₂)₂, or —P(O)(R₁₁)₂, wherein:

-   -   R₉ is a lower alkoxy, lower alkyl sulfanyl, or an alkoxyalkoxy;    -   R₁₁, for each occurrence, is independently, a lower alkyl; and    -   R₁₂, for each occurrence, is independently, H or a lower alkyl.

In another embodiment, the invention relates to compounds selected fromthe group consisting of:

-   3-Methyl-N-[5′-(pyridin-3-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide;-   3-Methyl-N-[5′-(isoxazol-5-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide;-   3-Methyl-N-[5′-(isoxazol-5-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide;-   3-Methyl-N-[5′-(3-methyl-isoxazol-5-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide;-   3-Methyl-N-[2′-methoxy-5′-(furan-2-yl)-biphenyl-4-yl]-isonicotinamide;-   3-Methyl-N-[5′-(thien-2-yl)-2′-methoxy-biphenyl-4-yl]-isonicotinamide;-   3-Methyl-N-[5′-([1,3,4]thiadiazol-2-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide;-   3-Fluoro-N-[5′-([1,3,4]thiadiazol-2-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide;    -   and pharmaceutically acceptable salts, solvates, clathrates, or        prodrugs thereof.

In another embodiment, the invention relates to compounds of formula(VIII):

-   -   or a pharmaceutically acceptable salt, solvate, clathrate, or        prodrug thereof, wherein:    -   X₁, L, Z, R₁, R₃, R₄ and n are defined as for formula (I).

In another embodiment of the compounds represented by formula (VII) or(VIII), n is 0.

In another embodiment of the compounds represented by formula (VII) or(VIII), X₁ is CH.

In another embodiment of the compounds represented by formula (VII) or(VIII), X₁ is N.

In another embodiment of the compounds represented by formula (VII) or(VIII), Z, for each occurrence, is independently, a lower alkyl, a loweralkoxy, a lower haloalkoxy, a halo, cyano, or haloalkyl and n is 1 or 2.

In another embodiment of the compounds represented by formula (VII) or(VIII), L is —NHC(O)—.

In another embodiment, the invention relates to compounds of formula(IX):

or a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof, wherein R₁, R₃, and R₄ are defined as above.

In another embodiment, the invention relates to compounds of formula(X):

or a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof, wherein R₁, R₃, and R₄ are defined as above.

In another embodiment of the compounds represented by formula (VII),(VIII), (IX) or (X), R₁ is a lower alkyl or a halo.

In another embodiment of the compounds represented by formula (VII),(VIII), (IX) or (X), R₃ is a lower alkyl, a lower alkoxy, a lower alkylsulfanyl, a lower alkylamino, a lower dialkylamino, or a halo.

In another embodiment of the compounds represented by formula (VII),(VIII), (IX) or (X), R₃ is a lower alkyl.

In another embodiment of the compounds represented by formula (VIII),(IX) or (X), R₄ is a bioisostere of an ester, amide, or carboxylic acid.

In another embodiment of the compounds represented by formula (VIII),(IX) or (X), R₄ is a 5-membered heteroaryl.

In another embodiment of the compounds represented by formula (VIII),(IX) or (X), R₄ is an optionally substituted oxazolyl, an optionallysubstituted thiazolyl, an optionally substituted 1H-tetrazolyl, anoptionally substituted 1H-imidazolyl, an optionally substituted[1,2,4]oxadiazolyl, or an optionally substituted 4H-[1,2,4]triazolyl.

In another embodiment of the compounds represented by formula (VIII),(IX) or (X), R₄ is a halo, —C(O)R₉, —S(O)_(p)R₁₁, —S(O)_(p)NR₅,—S(O)_(p)OR₅, —P(O)(OR₁₂)₂, or —P(O)(R₁₁)₂, wherein:

-   -   R₉ is a lower alkoxy, lower alkyl sulfanyl, or an alkoxyalkoxy;    -   R₁₁, for each occurrence, is independently, a lower alkyl; and    -   R₁₂, for each occurrence, is independently, H or a lower alkyl.

In another embodiment of the compounds represented by formula (VII),(VIII), (IX) or (X), R₁ is fluoro or methyl.

In another embodiment, the invention relates to compounds selected fromthe group consisting of:

-   3-Methyl-N-(2′-methyl-5′-oxazol-2-yl-biphenyl-4-yl)-isonicotinamide;-   3-Methyl-N-(2′-methyl-5′-thiazol-2-yl-biphenyl-4-yl)-isonicotinamide;-   3-Fluoro-N-(2′-methyl-5′-oxazol-2-yl-biphenyl-4-yl)-isonicotinamide;-   4′-[(3-Fluoro-pyridine-4-carbonyl)-amino]-6-methyl-biphenyl-3-carboxylic    acid methyl ester;-   3-Methyl-N-(2′-methyl-5′-thiazol-2-yl-biphenyl-4-yl)-isonicotin-amide;-   3-Methyl-N-(2′-chloro-5′-oxazol-2-yl-biphenyl-4-yl)-isonicotin-amide;-   3-Methyl-N-[4-(5-chloro-2-methoxy-pyridin-3-yl)-phenyl]-isonicotinamide;-   3-Methyl-N-[5′-(oxazol-5-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide;-   3-Methyl-N-[5′-(oxazol-5-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide;-   3-Methyl-N-[5′-(4-methyl-thiazol-2-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide;-   3-Methyl-N-[2′-methyl-5′-(1H-tetrazol-5-yl)-biphenyl-4-yl]-isonicotinamide;-   3-Methyl-N-[2′-methyl-5′-([1,3,4]oxadiazol-2-yl)-biphenyl-4-yl]-isonicotinamide;-   3-Methyl-N-[5′-(oxazol-2-yl)-2′-(N,N-dimethylamino)-biphenyl-4-yl]-isonicotinamide;-   3-Methyl-N-[2′-methyl-5′-(1-methyl-1H-tetrazol-5-yl)-biphenyl-4-yl]-isonicotinamide;-   3-Methyl-N-[5′-(oxazol-2-yl)-2′-methoxy-biphenyl-4-yl]-isonicotinamide;-   3-Fluoro-N-[5′-(thiazol-2-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide;-   3-Methyl-N-[5′-(thiazol-2-yl)-2′-chloro-biphenyl-4-yl]-isonicotinamide;-   3-Fluoro-N-[5′-(thiazol-2-yl)-2′-chloro-biphenyl-4-yl]-isonicotinamide;-   3-Fluoro-N-[2′-methyl-5′-(2-methyl-2H-[1,2,4]triazol-3-yl)-biphenyl-4-yl]-isonicotinamide;-   3-Methyl-N-[2′-methyl-5′-([1,3,4]oxadiazol-2-yl)-biphenyl-4-yl]-isonicotinamide;-   3-Fluoro-N-[2′-methyl-5′-([1,3,4]oxadiazol-2-yl)-biphenyl-4-yl]-isonicotinamide;-   3-Fluoro-N-[2′-methyl-5′-(4-methyl-5-methylsulfanyl-4H-[1,2,4]triazol-3-yl)-biphenyl-4-yl]-isonicotinamide;-   3-Methyl-N-[2′-methyl-5′-(4-methyl-5-methylsulfanyl-4H-[1,2,4]triazol-3-yl)-biphenyl-4-yl]-isonicotinamide;-   3-Fluoro-N-[2′-methyl-5′-(oxazol-4-yl)-biphenyl-4-yl]-isonicotinamide;-   3-Methyl-N-[2′-methyl-5′-(oxazol-4-yl)-biphenyl-4-yl]-isonicotinamide;-   3-Methyl-N-[2′-methyl-5′-(1H-tetrazol-5-yl)-biphenyl-4-yl]-isonicotinamide;    -   and pharmaceutically acceptable salts, solvates, clathrates, or        prodrugs thereof.

In another embodiment, the invention relates to compounds of formula(XI):

-   -   or a pharmaceutically acceptable salt, solvate, clathrate, or        prodrug thereof, wherein:    -   Z, R₃, R₁₈ and n are defined as for formula (I); and    -   Y is an optionally substituted 5- or 6-membered heteroaryl.

In one embodiment of the compounds represented by formula (XI), Y is anoptionally substituted 5-membered heteroaryl. For example, Y may be anoptionally substituted oxazolyl, an optionally substituted isoxazolyl,an optionally substituted pyrazolyl, an optionally substitutedthiazolyl, an optionally substituted pyrrolyl, an optionally substitutedfuranyl, an optionally substituted thienyl, an optionally substitutedthiadiazolyl, an optionally substituted triazolyl, an optionallysubstituted oxadiazolyl, or an optionally substituted tetrazolyl.Preferably, Y is an optionally substituted [1,2,3]thiadiazolyl.

In another embodiment, the invention relates to compounds of formula(XII):

-   -   or a pharmaceutically acceptable salt, solvate, clathrate, or        prodrug thereof, wherein:    -   Z, R₃, R₁₈ and n are defined as for formula (I); and    -   R₁₉ is H, a halo, an optionally substituted alkyl, an optionally        substituted alkoxy, or an optionally substituted alkyl sulfanyl.

In another embodiment of the compounds represented by formula (XII), R₁₉is a halo or a lower alkyl. Preferably, R₁₉ is a lower alkyl.

In another embodiment of the compounds represented by formula (X) or(XII), n is 0.

In another embodiment of the compounds represented by formula (X) or(XII), Z, for each occurrence, is independently, a lower alkyl, a loweralkoxy, a lower haloalkoxy, a halo, cyano, or haloalkyl and n is 1 or 2.

In another embodiment of the compounds represented by formula (X) or(XII), L is —NHC(O)—.

In another embodiment of the compounds represented by formula (XI) or(XI), R₃ is a lower alkyl, a lower alkoxy, a lower alkyl sulfanyl, alower alkylamino, a lower dialkylamino, or a halo.

In another embodiment of the compounds represented by formula (XI) or(XI), R₃ is a lower alkyl.

In one embodiment of the compounds represented by formula (XI), R₁₈ isan optionally substituted pyridinyl, an optionally substituted oxazolyl,an optionally substituted isoxazolyl, an optionally substitutedpyrazolyl, an optionally substituted thiazolyl, an optionallysubstituted pyrrolyl, an optionally substituted morpholinyl, anoptionally substituted furanyl, an optionally substituted thienyl, anoptionally substituted thiadiazolyl, an optionally substitutedtriazolyl, an optionally substituted oxadiazolyl, or an optionallysubstituted tetrazolyl. Preferably, R₁₈ is unsubstituted or substitutedwith one or more substituents selected from the group consisting of alower alkyl, halo, a lower haloalkyl, an amino, a lower dialkyl amino, alower alkyl amino, a lower alkoxy, and a lower alkyl sulfanyl.

In another embodiment of the compounds represented by formula (XI) or(XI), R₁₈ is a bioisostere of an ester, amide, or carboxylic acid.

In another embodiment of the compounds represented by formula (XI) or(XI), R₁₈ is a 5-membered heteroaryl.

In another embodiment of the compounds represented by formula (XI) or(XI), R₁₈ is an optionally substituted oxazolyl, an optionallysubstituted thiazolyl, an optionally substituted 1H-tetrazolyl, anoptionally substituted 1H-imidazolyl, an optionally substituted[1,2,4]oxadiazolyl, or an optionally substituted 4H-[1,2,4]triazolyl.

In another embodiment of the compounds represented by formula (XI) or(XI), R₁₈ is a halo, —C(O)R₉, —S(O)_(p)R₁₁, —S(O)_(p)NR₅, —S(O)_(p)OR₅,—P(O)(OR₁₂)₂, or —P(O)(R₁₁)₂, wherein:

-   -   R₉ is a lower alkoxy, lower alkyl sulfanyl, or an alkoxyalkoxy;    -   R₁₁, for each occurrence, is independently, a lower alkyl; and    -   R₁₂, for each occurrence, is independently, H or a lower alkyl.

In another embodiment, the invention relates to compounds selected fromthe group consisting of:

-   4-Methyl-[1,2,3]thiadazole-5-carboxylic acid    [2′-methyl-5′-(pyridin-3-yl)-biphenyl-4-yl]-amide;-   4-Methyl-[1,2,3]thiadazole-5-carboxylic acid    [2′-methyl-5′-(pyridin-2-yl)-biphenyl-4-yl]-amide;-   4-Methyl-[1,2,3]thiadiazole-5-carboxylic acid    [2′-methoxy-5′-(oxazol-5-yl)-biphenyl-4-yl]-amide;-   4-Methyl-[1,2,3]thiadazole-5-carboxylic acid    [2′-methyl-5′-(isoxazol-5-yl)-biphenyl-4-yl]-amide;-   4-Methyl-[1,2,3]thiadazole-5-carboxylic acid    [2′-methyl-5′-(thiazol-2-yl)-biphenyl-4-yl]-amide;    -   and pharmaceutically acceptable salts, solvates, clathrates, or        prodrugs thereof.

All of the features, specific embodiments and particular substituentsdisclosed herein may be combined in any combination. Each feature,embodiment or substituent disclosed in this specification may bereplaced by an alternative feature, embodiment or substituent servingthe same, equivalent, or similar purpose. In the case of chemicalcompounds, specific values for variables (e.g., values shown in theexemplary compounds disclosed herein) in any chemical formula disclosedherein can be combined in any combination resulting in a stablestructure. Furthermore, specific values (whether preferred or not) forsubstituents in one type of chemical structure may be combined withvalues for other substituents (whether preferred or not) in the same ordifferent type of chemical structure. Thus, unless expressly statedotherwise, each feature, embodiment or substituent disclosed is only anexample of a generic series of equivalent or similar features,embodiments or substituents.

In another embodiment, the invention relates to pharmaceuticalcompositions that comprise a compound of any one of formulas (I) through(XII), or Table 1, or a pharmaceutically acceptable salt, solvate,clathrate, or prodrug thereof, as an active ingredient, and apharmaceutically acceptable carrier or vehicle. The compositions areuseful for immunosuppression or to treat or prevent inflammatoryconditions, allergic conditions and immune disorders.

In another embodiment, the invention relates to methods forimmunosuppression or for treating or preventing inflammatory conditions,immune disorders, or allergic disorders in a patient in need thereofcomprising administering an effective amount of a compound representedby any one of formulas (I) through (XII), or Table 1, or apharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In another embodiment, the invention relates to methods forimmunosuppression or for treating or preventing inflammatory conditions,immune disorders, or allergic disorders in a patient in need thereofcomprising administering an effective amount of a pharmaceuticalcomposition that comprises a compound represented by any one of formulas(I) through (XII), or in or Table 1, or a pharmaceutically acceptablesalt, solvate, clathrate, or prodrug thereof.

In another embodiment, compounds of any one of formulas (I) through(XII), or Table 1, or a pharmaceutically acceptable salt, solvate,clathrate, or prodrug thereof, are particularly useful inhibiting immunecell (e.g., T-cells, B-cells, and/or mast cell) activation (e.g.,activation in response to an antigen) and/or T cell and/or B cellproliferation. Indicators of immune cell activation include secretion ofIL-2 by T cells, proliferation of T cells and/or B cells, and the like.The compounds of the invention inhibit IL-2 secretion by T-cells and/orB-cells. In one embodiment, a compound of any one of formulas (I)through (XII) or Table 1, inhibits immune cell activation and/or T celland/or B cell proliferation in a mammal (e.g., a human). In anotherembodiment, the compounds of the invention inhibit mast celldegranulation in response to an antigen.

In another embodiment, compounds of any one of formulas (I) through(XII), or Table 1, or a pharmaceutically acceptable salt, solvate,clathrate, or prodrug thereof, can inhibit the production of certaincytokines that regulate immune cell activation. For example, compoundsof any one of formulas (I) through (XII), or Table 1, or apharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof, can inhibit the production of IL-2, IL-4, IL-5, IL-13, GM-CSF,IFN-γ, TNF-α and combinations thereof. In one embodiment, a compound ofany one of formulas (I) through (XII), or Table 1, inhibits cytokineproduction in a mammal (e.g., a human).

In another embodiment, compounds of any one of formulas (I) through(XII), or Table 1, or a pharmaceutically acceptable salt, solvate,clathrate, or prodrug thereof, can modulate the activity of one or moreion channel involved in activation of immune cells, such as CRAG ionchannels. In one embodiment, a compound of any one of formulas (I)through (XII) or Table 1 can inhibit the influx of calcium ions into animmune cell (e.g., T cells and/or B cells and/or mast cells) byinhibiting the action of CRAC ion channels. The inhibition of the CRACchannel may be directed or indirect inhibition of the channel activity.In general, a decrease in I_(CRAC) current upon contacting a cell with acompound is one indicator that the compound inhibitions CRAC ionchannels. I_(CRAC) current can be measured, for example, using a patchclamp technique, which is described in more detail in the examplesbelow. In one embodiment, a compound of any one of formulas (I) through(XII) or Table 1 modulates an ion channel (e.g., CRAC channels) in amammal (e.g., a human).

Exemplary Compounds of the Invention

Exemplary compounds of the invention are depicted in Table 1 below.

TABLE 1 Compound No. Structure Chemical Name  1

4′-(2,6-Difluoro-benzoylamino)- 6-methyl-biphenyl-3-carboxylic acidmethyl ester  2

6-Chloro-4′-(2,6-difluoro- benzoylamino)-biphenyl-3- (carboxylic acid2-methoxyethyl ester)  3

2,6-Difluoro-N-(2′-methyl-5′- oxazol-2-yl-biphenyl-4-yl)- bezamide  4

4′-[(3,5-Difluoro-pyridine-4- cabonyl)-amino]-6-methyl-biphenyl-3-carboxylic acid methyl ester  5

N-[4-(5-Chloro-2-methoxy- pyridin-3-yl)-phenyl]-2,6-difluoro- benzamide 6

2,6-Difluoro-N-[2′-methyl-5′-(1H- tetrazol-5-yl)-biphenyl-4-yl]-benzamide  7

3-Methyl-N-(2′-methyl-5′-oxazol- 2-yl-biphenyl-4-yl)- isonicotinamide,hydrochloride  8

N-[4-(5-Chloro-2-methoxy- pyridin-3-yl)-phenyl]-2,6-difluoro- benzamide,hydrochloride  9

3-Methyl-N-(2′-methyl-5′-thiazol- 2-yl-biphenyl-4-yl)- isonicotinamide,HCl salt 10

3-Fluoro-N-(2′-methyl-5′-oxazol- 2-yl-biphenyl-4-yl)- isonicotinamide 11

3,5-Difluoro-N-(2′-methyl-5′- oxazol-2-yl-biphenyl-4-yl)-isonicotinamide 12

4′-[(3-Fluoro-pyridine-4- carbonyl)-amino]-6-methyl-biphenyl-3-carboxylic acid methyl ester 13

2,6-Difluoro-N-(2′-methoxy-5′- oxazol-2-yl-biphenyl-4-yl)- benzamide 14

3-Methyl-N-(2′-methyl-5′-thiazol- 2-yl-biphenyl-4-yl)-isonicotin- amide15

2,6-Difluoro-N-(2′-methyl-5′- thiazol-2-yl-biphenyl-4-yl)- benzamide 16

3-Methyl-N-(2′-chloro-5′-oxazol- 2-yl-biphenyl-4-yl)-isonicotin- amide17

3-Methyl-N-(2′-chloro-5′-oxazol- 2-yl-biphenyl-4-yl)-isonicotin- amide,hydrochloride 18

3-Methyl-N-[4-(5-chloro-2- methoxy-pyridin-3-yl)-phenyl]-isonicotinamide 19

3-Fluoro-N-[4-(5-chloro-2- methoxy-pyridin-3-yl)-phenyl]-isonicotinamide 20

3-Methyl-N-[5′-(pyridin-3-yl)-2′- methyl-biphenyl-4-yl]- isonicotinamide21

4-Methyl-[1,2,3]thiadazoie-5- carboxylic acid [2′-methyl-5′-(pyridin-3-yl)-biphenyl-4-yl]- amide 22

2,6-Difluoro-N-[2′-methyl-5′- (pyridine-3-yl)-biphenyl-4-yl]- benzamide23

2,6-Difluoro-N-[2′-methyl-5′- (pyridine-2-yl)-biphenyl-4-yl]- benzamide24

4-Methyl-[1,2,3]thiadazole-5- carboxylic acid [2′-methyl-5′-(pyridin-2-yl)-biphenyl-4-yl]- amide 25

4-Methyl-[1,2,3]thiadiazole-5- carboxylic acid [2′-methoxy-5′-(oxazol-5-yl)-biphenyl-4-yl]- amide 26

3-Methyl-N-[5′-(oxazol-5-yl)-2′- methyl-biphenyl-4-yl]- isonicotinamide27

2,6-Difluoro-N-[2′-methyl-5′- (oxazol-5-yl)-biphenyl-4-yl]- benzamide 28

3,5-Difluoro-N-[5′-(thiazol-2-yl)- 2′-methyl-biphenyl-4-yl]-isonicotinamide 29

3-Methyl-N-[5′-(oxazol-5-yl)-2′- methyl-biphenyl-4-yl]- isonicotinamide,HCl salt 30

2,6-Difluoro-N-[2′-methyl-5′- (pyridine-4-yl)-biphenyl-4-yl]- benzamide31

2,6-Difluoro-N-[2′-chloro-5′- (oxazol-2-yl)-biphenyl-4-yl]- benzamide 32

3-Methyl-N-[5′-(isoxazol-5-yl)-2′- methyl-biphenyl-4-yl]-isonicotinamide 33

4-Methyl-[1,2,3]thiadazole-5- carboxylic acid [2′-methyl-5′-(isoxazol-5-yl)-biphenyl-4-yl]- amide 34

3-Methyl-N-[5′-(isoxazol-5-yl)-2′- methyl-biphenyl-4-yl]-isonicotinamide, HCl salt 35

2,6-Difluoro-N-[2′-methyl-5′- (3-methyl-isoxazole-5-yl)-biphenyl-4-yl]-benzamide 36

3-Methyl-N-[5′-(3-methyl- isoxazol-5-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide 37

2,6-DifIuoro-N-[2′-methyl-5′- (3-methyl-1H-pyrazol-5-yl)-biphenyl-4-yl]-benzamide 38

2,6-Difluoro-N-[2′-methyl-5′- (4-methyl-thiazol-2-yl)-biphenyl-4-yl]-benzamide 39

2,6-Difluoro-N-[2′-methyl-5′- (4-trifluoromethyl-thiazol-2-yl)-biphenyl-4-yl]-benzamide 40

3-Methyl-N-[5′-(4-methyl- thiazol-2-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide 41

2,6-Difluoro-N-[2′-methyl-5′- (1H-pyrrol-2-yl)-biphenyl-4-yl]- benzamide42

2,6-Difluoro-N-[2′-methyl-5′- (5-oxo-4,5-dihydro-[1,2,4]-oxadiazol-3-yl)-biphenyl-4-yl]- benzamide 43

2,6-Difluoro-N-[2′-methyl-5′- (morpholino-4-yl)-biphenyl-4-yl]-benzamide 44

3-Methyl-N-[2′-methyl- 5′-(1H-tetrazol-5-yl)-biphenyl-4-yl]-isonicotinamide 45

3,5-Difluoro-N-[5′-(oxazol-2-yl)- 2′-chloro-biphenyl-4-yl]-isonicotinamide 46

3-Methyl-N-[2′-methyl- 5′-([1,3,4]oxadiazol-2-yl)-biphenyl-4-yl]-isonicotinamide, HCl salt 47

3-Methyl-N-[2′-methoxy-5′- (furan-2-yl)-biphenyl-4-yl]- isonicotinamide48

3-Methyl-N-[5′-(oxazol-2-yl)-2′- (N,N-dimethylamino)-biphenyl-4-yl]-isonicotinamide 49

3-Methyl-N-[2′-methyl-5′-(1- methyl-1H-tetrazol-5-yl)-biphenyl-4-yl]-isonicotinamide 50

3-Methyl-N-[5′-(oxazol-2-yl)-2′- methoxy-biphenyl-4-yl]- isonicotinamide51

3,5-Difluoro-N-[5′-(oxazol-2-yl)- 2′-methoxy-biphenyl-4-yl]-isonicotinamide 52

3-Methyl-N-[5′-(thien-2-yl)-2′- methoxy-biphenyl-4-yl]- isonicotinamide53

3,5-Difluoro-N-[5′- ([1,3,4]thiadiazol-2-yl)- 2′-methyl-biphenyl-4-yl]-isonicotinamide 54

3-Fluoro-N-[5′-(thiazol-2-yl)-2′- methyl-biphenyl-4-yl]- isonicotinamide55

3-Methyl-N-[5′-(thiazol-2-yl)-2′- chloro-biphenyl-4-yl]- isonicotinamide56

2,6-Difluoro-N-[2′-chloro-5′- (thiazol-2-yl)-biphenyl-4-yl]- benzamide57

3,5-Difluoro-N-[5′-(thiazol-2-yl)- 2′-chloro-biphenyl-4-yl]-isonicotinamide 58

3-Fluoro-N-[5′-(thiazol-2-yl)-2′- chloro-biphenyl-4-yl]- isonicotinamide59

2,6-Difluoro-N-[2′-methyl-5′- ([1,3,4]thiadiazol-2-yl)-biphenyl-4-yl]-benzamide 60

3-Methyl-N-[5′-([1,3,4]thiadiazol- 2-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide 61

3-Fluoro-N-[5′-([1,3,4]thiadiazol- 2-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide 62

2,6-Difluoro-N-[2′-methyl-5′- (5-amino-[1,3,4]thiadiazol-2-yl)-biphenyl-4-yl]-benzamide 63

2,6-Difluoro-N-{2′-methyl-5′- [5-(N,N-dimethylamino)-[1,3,4]thiadiazol-2-yl]-biphenyl- 4-yl}-N-methyl-benzamide 64

2,6-Difluoro-N-{2′-methyl-5′- [5-(N,N-dimethylamino)-[1,3,4]thiadiazol-2-yl]-biphenyl- 4-yl}-benzamide 65

2,6-Difluoro-N-[2′-methyl- 5′-(1H-tetrazol-5-yl)-biphenyl-4-yl]-benzamide, sodium salt 66

2,6-Difluoro-N-[2′-methyl- 5′-(2-methyl-2H-[1,2,4]triazol-3-yl)-biphenyl-4-yl]-benzamide 67

3-Fluoro-N-[2′-methyl- 5′-(2-methyl-2H-[1,2,4]triazol-3-yl)-biphenyl-4-yl]- isonicotinamide 68

3,5-Difluoro-N-[2′-methyl- 5′-([1,3,4]oxadiazol-2-yl)-biphenyl-4-yl]-isonicotinamide 69

2,6-Difluoro-N-[2′-methyl- 5′-([1,3,4]oxadiazol-2-yl)-biphenyl-4-yl]-benzamide 70

3-Methyl-N-[2′-methyl- 5′-([1,3,4]oxadiazol-2-yl)-biphenyl-4-yl]-isonicotinamide 71

3-Fluoro-N-[2′-methyl- 5′-([1,3,4]oxadiazol-2-yl)-biphenyl-4-yl]-isonicotinamide 72

2,6-Difluoro-N-[2′-methyl- 5′-(4-methyl-5-methylsulfanyl-4H-[1,2,4]triazol-3-yl)-biphenyl- 4-yl]-benzamide 73

3-Fluoro-N-[2′-methyl- 5′-(4-methyl-5-methylsulfanyl-4H-[1,2,4]triazol-3-yl)-biphenyl- 4-yl]-isonicotinamide 74

3-Methyl-N-[2′-methyl- 5′-(4-methyl-5-methylsulfanyl-4H-[1,2,4]triazol-3-yl)-biphenyl- 4-yl]-isonicotinamide 75

3,5-Difluoro-N-[2′-methyl- 5′-(4-methyl-5-methylsulfanyl-4H-[1,2,4]triazol-3-yl)-biphenyl- 4-yl]-isonicotinamide 76

4-Methyl-[1,2,3]thiadazole-5- carboxylic acid [2′-methyl-5′-(thiazol-2-yl)-biphenyl-4-yl]- amide 77

2,6-Difluoro-N-[2′-methyl- 5′-(oxazol-4-yl)-biphenyl- 4-yl]-benzamide 78

3-Fluoro-N-[2′-methyl- 5′-(oxazol-4-yl)-biphenyl- 4-yl]-isonicotinamide79

3-Methyl-N-[2′-methyl- 5′-(oxazol-4-yl)-biphenyl- 4-yl]-isonicotinamide80

3-Methyl-N-[2′-methyl- 5′-(1H-tetrazol-5-yl)-biphenyl-4-yl]-isonicotinamide, sodium salt 81

2,6-Difluoro-N-[2′-methyl- 5′-(1H-tetrazol-5-yl)-biphenyl-4-yl]-benzamide, HCl salt 82

2,6-Difluoro-N-[2′-methyl- 5′-(1-methyl-1H-tetrazol-5-yl)-biphenyl-4-yl]-benzamide 83

2,6-Difluoro-N-[2′-methyl- 5′-(2-methyl-2H-tetrazol-5-yl)-biphenyl-4-yl]-benzamide

Mechanism of Action

Activation of T-lymphocytes in response to an antigen is dependent oncalcium ion oscillations. Calcium ion oscillations in T-lymphocytes aretriggered through stimulation of the T-cell antigen receptor, andinvolve calcium ion influx through the stored-operatedCa²⁺-release-activated Ca²⁺ (CRAC) channel. Although the molecularstructure of the CRAC ion channel has not been identified, a detailedelectrophysiological profile of the channel exists. Thus, inhibition ofCRAC ion channels can be measured by measuring inhibition of theI_(CRAC) current. Calcium ion oscillations in T-cells have beenimplicated in the activation of several transcription factors (e.g.,NFAT, Oct/Oap and NFκB) which are critical for T-cell activation (Lewis,Biochemical Society Transactions (2003), 31:925-929, the entireteachings of which are incorporated herein by reference). Withoutwishing to be bound by any theory, it is believed that because thecompounds of the invention inhibit the activity of CRAC ion channels,they inhibit immune cell activation.

Methods of Treatment and Prevention

In accordance with the invention, an effective amount of a compound ofany one of formulas (I) through (XII) or Table 1, or a pharmaceuticallyacceptable salt, solvate, clathrate, and prodrug thereof, or apharmaceutical composition comprising a compound of any one of formulas(I) through (XII) or Table 1, or a pharmaceutically acceptable salt,solvate, clathrate, and prodrug thereof, is administered to a patient inneed of immunosuppression or in need of treatment or prevention of aninflammatory condition, an immune disorder, or an allergic disorder.Such patients may be treatment naïve or may experience partial or noresponse to conventional therapies.

Responsiveness of a particular inflammatory condition, immune disorder,or allergic disorder in a subject can be measured directly (e.g.,measuring blood levels of inflammatory cytokines (such as IL-2, IL-4,IL-5, IL-13, GM-CSF, TNF-α, IFN-γ and the like) after administration ofa compound of this invention), or can be inferred based on anunderstanding of disease etiology and progression. The compounds of anyone of formulas (I) through (XII), or Table 1, or pharmaceuticallyacceptable salts, solvates, clathrates, and prodrugs thereof can beassayed in vitro or in vivo, for the desired therapeutic or prophylacticactivity, prior to use in humans. For example, known animal models ofinflammatory conditions, immune disorders, or allergic disorders can beused to demonstrate the safety and efficacy of compounds of thisinvention.

Synthesis of Compounds of the Invention

In general, compounds of the invention that have an amide linker can beprepared via an amide coupling reaction, followed by a Suzuki couplingreaction (see Scheme A).

Alternatively, the Suzuki coupling reaction can be done first, followedby an amide coupling reaction (see Scheme B).

The amide coupling reaction can be accomplished by contacting an acidchloride with an amine in the presence of a base as shown in Scheme A orby contacting a carboxylic acid with an amine in the presence of a andialkylcarbodiimide, such as1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), as shown in SchemeB.

Compounds that have an amide group in the reverse direction (i.e., thecarbonyl portion of the amide is attached to the biphenyl orpyridinyl-phenyl group) can be synthesized by analogous methods as thoseshown in Schemes A and B.

In general methods for preparing compounds in which L is —NRCH₂—,—CH₂NR—, —C(O)—, —OC(O)—, —C(O)O—, —C(S)—, —NR—C(S)—, or —C(S)—NR— areknown in the art and can be found, for example, in March, AdvancedOrganic Chemistry, third edition, (1985), John Wiley & Sons, the entireteachings of which are incorporated herein by reference. Examples ofsuch methods are described briefly below.

Compounds in which L is —NRCH₂— or —CH₂NR— can be prepared fromcompounds that have amide linkers by reducing the amide group withsodium borohydride. Typically, the reaction is carried out in an alcoholsolvent, such as ethanol, and the reaction is heated.

Compounds in which L is —C(O)O— or —OC(O)— can be prepared by a methodanalogous to the amide coupling reaction except that the —NH₂ group isreplaced with a hydroxyl group.

Compounds in which L is —C(S)NR— or —NRC(S)— can be prepared by treatingcompounds that have an amide linker with2,4-bis(4-methoxyphenyl)-1,3,2,4-dithiadiphosphetane-2,4-disulfied (seePedersen, et al., Bul. Soc. Chim. Beiges (1978), 87:223, the entireteachings of which are incorporated herein by reference). Compounds inwhich L is —C(S)— can be prepared from compounds that have a —C(O)—linker by a similar method.

Methods of preparing the compounds of the invention are described inmore detail below in the example. Additional methods for preparingcompounds of the invention can be found in U.S. patent application Ser.No. 10/897,681, filed on Jul. 22, 2004, the entire teachings of whichare incorporated herein by reference.

Pharmaceutical Compositions and Dosage Forms

Pharmaceutical compositions and dosage forms of the invention compriseone or more active ingredients in relative amounts and formulated insuch a way that a given pharmaceutical composition or dosage form can beused for immunosuppression or to treat or prevent inflammatoryconditions, immune disorders, and allergic disorders. Preferredpharmaceutical compositions and dosage forms comprise a compound of anyone of formulas (I) through (XII), or Table 1, or a pharmaceuticallyacceptable prodrug, salt, solvate, or clathrate thereof, optionally incombination with one or more additional active agents.

Single unit dosage forms of the invention are suitable for oral, mucosal(e.g., nasal, sublingual, vaginal, buccal, or rectal), parenteral (e.g.,subcutaneous, intravenous, bolus injection, intramuscular, orintraarterial), or transdermal administration to a patient. Examples ofdosage forms include, but are not limited to: tablets; caplets;capsules, such as soft elastic gelatin capsules; cachets; troches;lozenges; dispersions; suppositories; ointments; cataplasms (poultices);pastes; powders; dressings; creams; plasters; solutions; patches;aerosols (e.g., nasal sprays or inhalers); gels; liquid dosage formssuitable for oral or mucosal administration to a patient, includingsuspensions (e.g., aqueous or non-aqueous liquid suspensions,oil-in-water emulsions, or a water-in-oil liquid emulsions), solutions,and elixirs; liquid dosage forms suitable for parenteral administrationto a patient; and sterile solids (e.g., crystalline or amorphous solids)that can be reconstituted to provide liquid dosage forms suitable forparenteral administration to a patient.

The composition, shape, and type of dosage forms of the invention willtypically vary depending on their use. For example, a dosage formsuitable for mucosal administration may contain a smaller amount ofactive ingredient(s) than an oral dosage form used to treat the sameindication. This aspect of the invention will be readily apparent tothose skilled in the art. See, e.g., Remington's Pharmaceutical Sciences(1990) 18th ed., Mack Publishing, Easton Pa.

Typical pharmaceutical compositions and dosage forms comprise one ormore excipients. Suitable excipients are well known to those skilled inthe art of pharmacy, and non-limiting examples of suitable excipientsare provided herein. Whether a particular excipient is suitable forincorporation into a pharmaceutical composition or dosage form dependson a variety of factors well known in the art including, but not limitedto, the way in which the dosage form will be administered to a patient.For example, oral dosage forms such as tablets may contain excipientsnot suited for use in parenteral dosage forms.

The suitability of a particular excipient may also depend on thespecific active ingredients in the dosage form. For example, thedecomposition of some active ingredients can be accelerated by someexcipients such as lactose, or when exposed to water. Active ingredientsthat comprise primary or secondary amines (e.g., N-desmethylvenlafaxineand N,N-didesmethylvenlafaxine) are particularly susceptible to suchaccelerated decomposition. Consequently, this invention encompassespharmaceutical compositions and dosage forms that contain little, ifany, lactose. As used herein, the term “lactose-free” means that theamount of lactose present, if any, is insufficient to substantiallyincrease the degradation rate of an active ingredient. Lactose-freecompositions of the invention can comprise excipients that are wellknown in the art and are listed, for example, in the U.S. Pharmocopia(USP)SP (XXI)/NF (XVI). In general, lactose-free compositions compriseactive ingredients, a binder/filler, and a lubricant in pharmaceuticallycompatible and pharmaceutically acceptable amounts. Preferredlactose-free dosage forms comprise active ingredients, microcrystallinecellulose, pre-gelatinized starch, and magnesium stearate.

This invention further encompasses anhydrous pharmaceutical compositionsand dosage forms comprising active ingredients, since water canfacilitate the degradation of some compounds. For example, the additionof water (e.g., 5%) is widely accepted in the pharmaceutical arts as ameans of simulating long-term storage in order to determinecharacteristics such as shelf-life or the stability of formulations overtime. See, e.g., Jens T. Carstensen (1995) Drug Stability: Principles &Practice, 2d. Ed., Marcel Dekker, NY, N.Y., 379-80. In effect, water andheat accelerate the decomposition of some compounds. Thus, the effect ofwater on a formulation can be of great significance since moistureand/or humidity are commonly encountered during manufacture, handling,packaging, storage, shipment, and use of formulations.

Anhydrous pharmaceutical compositions and dosage forms of the inventioncan be prepared using anhydrous or low moisture containing ingredientsand low moisture or low humidity conditions. Pharmaceutical compositionsand dosage forms that comprise lactose and at least one activeingredient that comprises a primary or secondary amine are preferablyanhydrous if substantial contact with moisture and/or humidity duringmanufacturing, packaging, and/or storage is expected.

An anhydrous pharmaceutical composition should be prepared and storedsuch that its anhydrous nature is maintained. Accordingly, anhydrouscompositions are preferably packaged using materials known to preventexposure to water such that they can be included in suitable formularykits. Examples of suitable packaging include, but are not limited to,hermetically sealed foils, plastics, unit dose containers (e.g., vials),blister packs, and strip packs.

The invention further encompasses pharmaceutical compositions and dosageforms that comprise one or more compounds that reduce the rate by whichan active ingredient will decompose. Such compounds, which are referredto herein as “stabilizer” include, but are not limited to, antioxidantssuch as ascorbic acid, pH buffers, or salt buffers.

Like the amounts and types of excipients, the amounts and specific typesof active ingredients in a dosage form may differ depending on factorssuch as, but not limited to, the route by which it is to be administeredto patients. However, typical dosage forms of the invention comprise acompound of any one of formulas (I) through (XII), or Table 1, or apharmaceutically acceptable salt, solvate, clathrate, or prodrug thereofin an amount of from about 1 mg to about 1000 mg, preferably in anamount of from about 50 mg to about 500 mg, and most preferably in anamount of from about 75 mg to about 350 mg. The typical total dailydosage of a compound of any one of formulas (I) through (XII), or Table1, or a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof can range from about 1 mg to about 5000 mg per day, preferablyin an amount from about 50 mg to about 1500 mg per day, more preferablyfrom about 75 mg to about 1000 mg per day. It is within the skill of theart to determine the appropriate dose and dosage form for a givenpatient.

Oral Dosage Forms

Pharmaceutical compositions of the invention that are suitable for oraladministration can be presented as discrete dosage forms, such as, butare not limited to, tablets (e.g., chewable tablets), caplets, capsules,and liquids (e.g., flavored syrups). Such dosage forms containpredetermined amounts of active ingredients, and may be prepared bymethods of pharmacy well known to those skilled in the art. Seegenerally, Remington's Pharmaceutical Sciences (1990) 18th ed., MackPublishing, Easton Pa.

Typical oral dosage forms of the invention are prepared by combining theactive ingredient(s) in an admixture with at least one excipientaccording to conventional pharmaceutical compounding techniques.Excipients can take a wide variety of forms depending on the form ofpreparation desired for administration. For example, excipients suitablefor use in oral liquid or aerosol dosage forms include, but are notlimited to, water, glycols, oils, alcohols, flavoring agents,preservatives, and coloring agents. Examples of excipients suitable foruse in solid oral dosage forms (e.g., powders, tablets, capsules, andcaplets) include, but are not limited to, starches, sugars,micro-crystalline cellulose, diluents, granulating agents, lubricants,binders, and disintegrating agents.

Because of their ease of administration, tablets and capsules representthe most advantageous oral dosage unit forms, in which case solidexcipients are employed. If desired, tablets can be coated by standardaqueous or nonaqueous techniques. Such dosage forms can be prepared byany of the methods of pharmacy. In general, pharmaceutical compositionsand dosage forms are prepared by uniformly and intimately admixing theactive ingredients with liquid carriers, finely divided solid carriers,or both, and then shaping the product into the desired presentation ifnecessary.

For example, a tablet can be prepared by compression or molding.Compressed tablets can be prepared by compressing in a suitable machinethe active ingredients in a free-flowing form such as powder orgranules, optionally mixed with an excipient. Molded tablets can be madeby molding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

Examples of excipients that can be used in oral dosage forms of theinvention include, but are not limited to, binders, fillers,disintegrants, and lubricants. Binders suitable for use inpharmaceutical compositions and dosage forms include, but are notlimited to, corn starch, potato starch, or other starches, gelatin,natural and synthetic gums such as acacia, sodium alginate, alginicacid, other alginates, powdered tragacanth, guar gum, cellulose and itsderivatives (e.g., ethyl cellulose, cellulose acetate, carboxymethylcellulose calcium, sodium carboxymethyl cellulose), polyvinylpyrrolidone, methyl cellulose, pre-gelatinized starch, hydroxypropylmethyl cellulose, (e.g., Nos. 2208, 2906, 2910), microcrystallinecellulose, and mixtures thereof.

Suitable forms of microcrystalline cellulose include, but are notlimited to, the materials sold as AVICEL-PH-101, AVICEL-PH-103 AVICELRC-581, AVICEL-PH-105 (available from FMC Corporation, American ViscoseDivision, Avicel Sales, Marcus Hook, Pa.), and mixtures thereof. Onespecific binder is a mixture of microcrystalline cellulose and sodiumcarboxymethyl cellulose sold as AVICEL RC-581. Suitable anhydrous or lowmoisture excipients or additives include AVICEL-PH-103J and Starch 1500LM.

Examples of fillers suitable for use in the pharmaceutical compositionsand dosage forms disclosed herein include, but are not limited to, talc,calcium carbonate (e.g., granules or powder), microcrystallinecellulose, powdered cellulose, dextrates, kaolin, mannitol, silicicacid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof.The binder or filler in pharmaceutical compositions of the invention istypically present in from about 50 to about 99 weight percent of thepharmaceutical composition or dosage form.

Disintegrants are used in the compositions of the invention to providetablets that disintegrate when exposed to an aqueous environment.Tablets that contain too much disintegrant may disintegrate in storage,while those that contain too little may not disintegrate at a desiredrate or under the desired conditions. Thus, a sufficient amount ofdisintegrant that is neither too much nor too little to detrimentallyalter the release of the active ingredients should be used to form solidoral dosage forms of the invention. The amount of disintegrant usedvaries based upon the type of formulation, and is readily discernible tothose of ordinary skill in the art. Typical pharmaceutical compositionscomprise from about 0.5 to about 15 weight percent of disintegrant,preferably from about 1 to about 5 weight percent of disintegrant.

Disintegrants that can be used in pharmaceutical compositions and dosageforms of the invention include, but are not limited to, agar-agar,alginic acid, calcium carbonate, microcrystalline cellulose,croscarmellose sodium, crospovidone, polacrilin potassium, sodium starchglycolate, potato or tapioca starch, other starches, pre-gelatinizedstarch, other starches, clays, other algins, other celluloses, gums, andmixtures thereof.

Lubricants that can be used in pharmaceutical compositions and dosageforms of the invention include, but are not limited to, calciumstearate, magnesium stearate, mineral oil, light mineral oil, glycerin,sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid,sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanutoil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, andsoybean oil), zinc stearate, ethyl oleate, ethyl laureate, agar, andmixtures thereof. Additional lubricants include, for example, a syloidsilica gel (AEROSIL 200, manufactured by W.R. Grace Co. of Baltimore,Md.), a coagulated aerosol of synthetic silica (marketed by Degussa Co.of Plano, Tex.), CAB-O-SIL (a pyrogenic silicon dioxide product sold byCabot Co. of Boston, Mass.), and mixtures thereof. If used at all,lubricants are typically used in an amount of less than about 1 weightpercent of the pharmaceutical compositions or dosage forms into whichthey are incorporated.

Controlled Release Dosage Forms

Active ingredients of the invention can be administered by controlledrelease means or by delivery devices that are well known to those ofordinary skill in the art. Examples include, but are not limited to,those described in U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809;3,598,123; and 4,008,719, 5,674,533, 5,059,595, 5,591,767, 5,120,548,5,073,543, 5,639,476, 5,354,556, and 5,733,566, each of which isincorporated herein by reference. Such dosage forms can be used toprovide slow or controlled-release of one or more active ingredientsusing, for example, hydropropylmethyl cellulose, other polymer matrices,gels, permeable membranes, osmotic systems, multilayer coatings,microparticles, liposomes, microspheres, or a combination thereof toprovide the desired release profile in varying proportions. Suitablecontrolled-release formulations known to those of ordinary skill in theart, including those described herein, can be readily selected for usewith the active ingredients of the invention. The invention thusencompasses single unit dosage forms suitable for oral administrationsuch as, but not limited to, tablets, capsules, gelcaps, and capletsthat are adapted for controlled-release.

All controlled-release pharmaceutical products have a common goal ofimproving drug therapy over that achieved by their non-controlledcounterparts. Ideally, the use of an optimally designedcontrolled-release preparation in medical treatment is characterized bya minimum of drug substance being employed to cure or control thecondition in a minimum amount of time. Advantages of controlled-releaseformulations include extended activity of the drug, reduced dosagefrequency, and increased patient compliance. In addition,controlled-release formulations can be used to affect the time of onsetof action or other characteristics, such as blood levels of the drug,and can thus affect the occurrence of side (e.g., adverse) effects.

Most controlled-release formulations are designed to initially releasean amount of drug (active ingredient) that promptly produces the desiredtherapeutic effect, and gradually and continually release of otheramounts of drug to maintain this level of therapeutic or prophylacticeffect over an extended period of time. In order to maintain thisconstant level of drug in the body, the drug must be released from thedosage form at a rate that will replace the amount of drug beingmetabolized and excreted from the body. Controlled-release of an activeingredient can be stimulated by various conditions including, but notlimited to, pH, temperature, enzymes, water, or other physiologicalconditions or compounds.

A particular extended release formulation of this invention comprises atherapeutically or prophylactically effective amount of a compound offormulas (I) through (XII), or Table 1, or a pharmaceutically acceptablesalt, solvate, hydrate, clathrate, or prodrug thereof, in spheroidswhich further comprise microcrystalline cellulose and, optionally,hydroxypropylmethyl-cellulose coated with a mixture of ethyl celluloseand hydroxypropylmethylcellulose. Such extended release formulations canbe prepared according to U.S. Pat. No. 6,274,171, the entire teachingsof which are incorporated herein by reference.

A specific controlled-release formulation of this invention comprisesfrom about 6% to about 40% a compound of any one of formulas (I) through(XII), or Table 1 by weight, about 50% to about 94% microcrystallinecellulose, NF, by weight, and optionally from about 0.25% to about 1% byweight of hydroxypropyl-methylcellulose, USP, wherein the spheroids arecoated with a film coating composition comprised of ethyl cellulose andhydroxypropylmethylcellulose.

Parenteral Dosage Forms

Parenteral dosage forms can be administered to patients by variousroutes including, but not limited to, subcutaneous, intravenous(including bolus injection), intramuscular, and intraarterial. Becausetheir administration typically bypasses patients' natural defensesagainst contaminants, parenteral dosage forms are preferably sterile orcapable of being sterilized prior to administration to a patient.Examples of parenteral dosage forms include, but are not limited to,solutions ready for injection, dry products ready to be dissolved orsuspended in a pharmaceutically acceptable vehicle for injection,suspensions ready for injection, and emulsions.

Suitable vehicles that can be used to provide parenteral dosage forms ofthe invention are well known to those skilled in the art. Examplesinclude, but are not limited to: Water for Injection USP; aqueousvehicles such as, but not limited to, Sodium Chloride Injection,Ringer's Injection, Dextrose Injection, Dextrose and Sodium ChlorideInjection, and Lactated Ringer's Injection; water-miscible vehicles suchas, but not limited to, ethyl alcohol, polyethylene glycol, andpolypropylene glycol; and non-aqueous vehicles such as, but not limitedto, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate,isopropyl myristate, and benzyl benzoate.

Compounds that increase the solubility of one or more of the activeingredients disclosed herein can also be incorporated into theparenteral dosage forms of the invention.

Transdermal, Topical, and Mucosal Dosage Forms

Transdermal, topical, and mucosal dosage forms of the invention include,but are not limited to, ophthalmic solutions, sprays, aerosols, creams,lotions, ointments, gels, solutions, emulsions, suspensions, or otherforms known to one of skill in the art. See, e.g., Remington'sPharmaceutical Sciences (1980 & 1990) 16th and 18th eds., MackPublishing, Easton Pa. and Introduction to Pharmaceutical Dosage Forms(1985) 4th ed., Lea & Febiger, Philadelphia. Dosage forms suitable fortreating mucosal tissues within the oral cavity can be formulated asmouthwashes or as oral gels. Further, transdermal dosage forms include“reservoir type” or “matrix type” patches, which can be applied to theskin and worn for a specific period of time to permit the penetration ofa desired amount of active ingredients.

Suitable excipients (e.g., carriers and diluents) and other materialsthat can be used to provide transdermal, topical, and mucosal dosageforms encompassed by this invention are well known to those skilled inthe pharmaceutical arts, and depend on the particular tissue to which agiven pharmaceutical composition or dosage form will be applied. Withthat fact in mind, typical excipients include, but are not limited to,water, acetone, ethanol, ethylene glycol, propylene glycol,butane-1,3-diol, isopropyl myristate, isopropyl palmitate, mineral oil,and mixtures thereof to form lotions, tinctures, creams, emulsions, gelsor ointments, which are non-toxic and pharmaceutically acceptable.Moisturizers or humectants can also be added to pharmaceuticalcompositions and dosage forms if desired. Examples of such additionalingredients are well known in the art. See, e.g., Remington'sPharmaceutical Sciences (1980 & 1990) 16th and 18th eds., MackPublishing, Easton Pa.

Depending on the specific tissue to be treated, additional componentsmay be used prior to, in conjunction with, or subsequent to treatmentwith active ingredients of the invention. For example, penetrationenhancers can be used to assist in delivering the active ingredients tothe tissue. Suitable penetration enhancers include, but are not limitedto: acetone; various alcohols such as ethanol, oleyl, andtetrahydrofuryl; alkyl sulfoxides such as dimethyl sulfoxide; dimethylacetamide; dimethyl formamide; polyethylene glycol; pyrrolidones such aspolyvinylpyrrolidone; Kollidon grades (Povidone, Polyvidone); urea; andvarious water-soluble or insoluble sugar esters such as Tween 80(polysorbate 80) and Span 60 (sorbitan monostearate).

The pH of a pharmaceutical composition or dosage form, or of the tissueto which the pharmaceutical composition or dosage form is applied, mayalso be adjusted to improve delivery of one or more active ingredients.Similarly, the polarity of a solvent carrier, its ionic strength, ortonicity can be adjusted to improve delivery. Compounds such asstearates can also be added to pharmaceutical compositions or dosageforms to advantageously alter the hydrophilicity or lipophilicity of oneor more active ingredients so as to improve delivery. In this regard,stearates can serve as a lipid vehicle for the formulation, as anemulsifying agent or surfactant, and as a delivery-enhancing orpenetration-enhancing agent. Different salts, hydrates or solvates ofthe active ingredients can be used to further adjust the properties ofthe resulting composition.

Combination Therapy

The methods for immunosuppression or for treating or preventinginflammatory conditions and immune disorders in a patient in needthereof can further comprise administering to the patient beingadministered a compound of this invention, an effective amount of one ormore other active agents. Such active agents may include those usedconventionally for immunosuppression or for inflammatory conditions orimmune disorders. These other active agents may also be those thatprovide other benefits when administered in combination with thecompounds of this invention. For example, other therapeutic agents mayinclude, without limitation, steroids, non-steroidal anti-inflammatoryagents, antihistamines, analgesics, immunosuppressive agents andsuitable mixtures thereof. In such combination therapy treatment, boththe compounds of this invention and the other drug agent(s) areadministered to a subject (e.g., humans, male or female) by conventionalmethods. The agents may be administered in a single dosage form or inseparate dosage forms. Effective amounts of the other therapeutic agentsand dosage forms are well known to those skilled in the art. It is wellwithin the skilled artisan's purview to determine the other therapeuticagent's optimal effective-amount range.

In one embodiment of the invention where another therapeutic agent isadministered to a subject, the effective amount of the compound of thisinvention is less than its effective amount when the other therapeuticagent is not administered. In another embodiment, the effective amountof the conventional agent is less than its effective amount when thecompound of this invention is not administered. In this way, undesiredside effects associated with high doses of either agent may beminimized. Other potential advantages (including without limitationimproved dosing regimens and/or reduced drug cost) will be apparent tothose of skill in the art.

In one embodiment relating to autoimmune and inflammatory conditions,the other therapeutic agent may be a steroid or a non-steroidalanti-inflammatory agent. Particularly useful non-steroidalanti-inflammatory agents, include, but are not limited to, aspirin,ibuprofen, diclofenac, naproxen, benoxaprofen, flurbiprofen, fenoprofen,flubufen, ketoprofen, indoprofen, piroprofen, carprofen, oxaprozin,pramoprofen, muroprofen, trioxaprofen, suprofen, aminoprofen,tiaprofenic acid, fluprofen, bucloxic acid, indomethacin, sulindac,tolmetin, zomepirac, tiopinac, zidometacin, acemetacin, fentiazac,clidanac, oxpinac, mefenamic acid, meclofenamic acid, flufenamic acid,niflumic acid, tolfenamic acid, diflurisal, flufenisal, piroxicam,sudoxicam, isoxicam; salicylic acid derivatives, including aspirin,sodium salicylate, choline magnesium trisalicylate, salsalate,diflunisal, salicylsalicylic acid, sulfasalazine, and olsalazin;para-aminophennol derivatives including acetaminophen and phenacetin;indole and indene acetic acids, including indomethacin, sulindac, andetodolac; heteroaryl acetic acids, including tolmetin, diclofenac, andketorolac; anthranilic acids (fenamates), including mefenamic acid, andmeclofenamic acid; enolic acids, including oxicams (piroxicam,tenoxicam), and pyrazolidinediones (phenylbutazone, oxyphenthartazone);and alkanones, including nabumetone and pharmaceutically acceptablesalts thereof and mixtures thereof. For a more detailed description ofthe NSAIDs, see Paul A. Insel, Analgesic-Antipyretic andAntiinflammatory Agents and Drugs Employed in the Treatment of Gout, inGoodman & Gilman's The Pharmacological Basis of Therapeutics 617-57(Perry B. Molinhoff and Raymond W. Ruddon eds., 9^(th) ed 1996) and GlenR. Hanson, Analgesic, Antipyretic and Anti-Inflammatory Drugs inRemington: The Science and Practice of Pharmacy Vol II 1196-1221 (A.R.Gennaro ed. 19th ed. 1995) which are hereby incorporated by reference intheir entireties.

Of particular relevance to allergic disorders, the other therapeuticagent may be an anthihistamine. Useful antihistamines include, but arenot limited to, loratadine, cetirizine, fexofenadine, desloratadine,diphenhydramine, chlorpheniramine, chlorcyclizine, pyrilamine,promethazine, terfenadine, doxepin, carbinoxamine, clemastine,tripelennamine, brompheniramine, hydroxyzine, cyclizine, meclizine,cyproheptadine, phenindamine, acrivastine, azelastine, levocabastine,and mixtures thereof. For a more detailed description ofanthihistamines, see Goodman & Gilman's The Pharmacological Basis ofTherapeutics (2001) 651-57, 10^(th) ed).

Immunosuppressive agents include glucocorticoids, corticosteroids (suchas Prednisone or Solumedrol), T cell blockers (such as cyclosporin A andFK506), purine analogs (such as azathioprine (Imuran)), pyrimidineanalogs (such as cytosine arabinoside), alkylating agents (such asnitrogen mustard, phenylalanine mustard, buslfan, and cyclophosphamide),folic acid antagonsists (such as aminopterin and methotrexate),antibiotics (such as rapamycin, actinomycin D, mitomycin C, puramycin,and chloramphenicol), human IgG, antilymphocyte globulin (ALG), andantibodies (such as anti-CD3 (OKT3), anti-CD4 (OKT4), anti-CD5,anti-CD7, anti-IL-2 receptor, anti-alpha/beta TCR, anti-ICAM-1,anti-CD20 (Rituxan), anti-IL-12 and antibodies to immunotoxins).

The foregoing and other useful combination therapies will be understoodand appreciated by those of skill in the art. Potential advantages ofsuch combination therapies include a different efficacy profile, theability to use less of each of the individual active ingredients tominimize toxic side effects, synergistic improvements in efficacy,improved ease of administration or use and/or reduced overall expense ofcompound preparation or formulation.

Other Embodiments

The compounds of this invention may be used as research tools (forexample, as a positive control for evaluating other potential CRACinhibitors, or IL-2, IL-4, IL-5, IL-13, GM-CSF, TNF-α, and/or INF-γinhibitors). These and other uses and embodiments of the compounds andcompositions of this invention will be apparent to those of ordinaryskill in the art.

The invention is further defined by reference to the following examplesdescribing in detail the preparation of compounds of the invention. Itwill be apparent to those skilled in the art that many modifications,both to materials and methods, may be practiced without departing fromthe purpose and interest of this invention. The following examples areset forth to assist in understanding the invention and should not beconstrued as specifically limiting the invention described and claimedherein. Such variations of the invention, including the substitution ofall equivalents now known or later developed, which would be within thepurview of those skilled in the art, and changes in formulation or minorchanges in experimental design, are to be considered to fall within thescope of the invention incorporated herein.

EXAMPLES Experimental Rationale

Without wishing to be bound by theory, it is believed that the compoundsof this invention inhibit CRAC ion channels, thereby inhibitingproduction of IL-2 and other key cytokines involved with inflammatoryand immune responses. The examples that follow demonstrate theseproperties.

Materials and General Methods

Reagents and solvents used below can be obtained from commercial sourcessuch as Aldrich Chemical Co. (Milwaukee, Wis., USA). ¹H-NMR and ¹³C-NMRspectra were recorded on a Varian 300 MHz NMR spectrometer. Significantpeaks are tabulated in the order: δ (ppm): chemical shift, multiplicity(s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; br s,broad singlet), coupling constant(s) in Hertz (Hz) and number ofprotons.

Patch clamp experiments were performed in the tight-seal whole-cellconfiguration at 21-25° C. High resolution current recordings wereacquired by a computer-based patch clamp amplifier system (EPC-9, HEKA,Lambrecht, Germany). Patch pipettes had resistances between 2-4 MΩ afterfilling with the standard intracellular solution. Immediately followingestablishment of the whole-cell configuration, voltage ramps of 50-200ms duration spanning the voltage range of −100 to +100 mV were deliveredat a rate of 0.5 Hz over a period of 300-400 seconds. All voltages werecorrected for a liquid junction potential of 10 mV between external andinternal solutions when using glutamate as the intracellular anion.Currents were filtered at 2.9 kHz and digitized at 10 μs intervals.Capacitive currents and series resistance were determined and correctedbefore each voltage ramp using the automatic capacitance compensation ofthe EPC-9. The low resolution temporal development of membrane currentswas assessed by extracting the current amplitude at −80 mV or +80 mVfrom individual ramp current records.

Example 1 Synthesis of Representative Exemplary Compounds of thisInvention Compound 1:4′-(2,6-difluoro-benzoylamino)-6-methyl-biphenyl-3-carboxylic acidmethyl ester

Step A:

To a stirred solution of4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenylamine (5.2 g, 24mmol), TEA (5 mL) in dry DCM (50 mL) at 0° C. was added2,6-difluoro-benzoyl chloride (3.0 mL, 24 mmol) dropwise. The mixturewas allowed to warm to room temperature over 2 h before it was washedwith water (2×100 mL) and dried. Removal of solvents gave2,6-difluoro-N-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-benzamide(8.4 g, 23 mmol) as white solid.

¹H-NMR (CDCl₃) δ (ppm) 7.8 (d, 2H, J=8), 7.7 (br, 1H), 7.6 (m, 2H), 7.4(m, 1H), 7.0 (t, 2H, J=9), 1.35 (s, 12H); ESMS clcd for C₁₉H₂₀BF₂NO₃:359.1. Found: 360.1 (M+H)⁺.

Step B:

A suspension of2,6-difluoro-N-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-benzamide(359 mg, 1 mmol), 3-bromo-4-methyl-benzoic acid methyl ester (228 mg, 1mmol), benzyl(chloro)bis(triphenylphosphine)palladium (38 mg, 0.05 mmol)and K₂CO₃ (690 mg, 5 mmol) in 1-methylpyrrolidinone (NMP) (5 ml) wasdegassed with vacuum and heated at 120° C. for 10 hr. After cooling downto room temperature, ethyl acetate (EtOAc) (200 ml) was added and themixture was washed with water (60 ml×3). The EtOAc was evaporated andthe residue was purified by column chromatography on silica gel(Hexanes:EtOAc) to give Compound 1 (286 mg, yield 75%).

¹H NMR (300 MHz, CDCl₃): 7.96-6.95 (m, 10H), 3.88 (s, 3H), 2.32 (s, 3H).ESMS calcd (C₂₂H₁₇F₂NO₃):381.12. found: 382.1 (M+H).

Compound 2:6-Chloro-4′-(2,6-difluoro-benzoylamino)-biphenyl-3-carboxylic acid2-methoxy-ethyl ester

Step A:

A mixture of2,6-difluoro-N-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-benzamide(0.35 g, 1 mmol), 4-chloro-3-iodo-benzoic acid (c) (0.28 g, 1 mmol),PdCl₂(PPh₃)₂ (80 mg, 0.1 mmol), and K₂CO₃ (0.14 g, 1 mmol) in NMP (4 mL)was stirred at 130° C. under nitrogen for 24 h. After being cooled, themixture was poured into ice-water (50 mL). The resultant precipitationwas collected by filtration, then washed with water. The solid materialwas dried and dissolved in DCM, undissolved material was filtered off.0.12 g (30%) of pure product,6-chloro-4′-(2,6-difluoro-benzoylamino)-biphenyl-3-carboxylic acid, wasobtained by silica gel chromatography (hexane/EtOAc to EtOAc/MeOH).

¹H-NMR (CDCl₃) δ ppm 7.30 (t, 2H, J=7), 7.40-8.15 (m, 9H); ESMS calcdfor C₂₀H₁₂ClF₂NO₃: 387.0. found: 388.0 (M+H).

Step B:

To a stirred solution of6-chloro-4′-(2,6-difluoro-benzoylamino)-biphenyl-3-carboxylic acid (13mg, 36 umol), 2-methoxy-ethanol (2.6 mg, 36 umol), andtriphenylphosphine (PPh₃) (10 mg, 38 umol) in dry THF (0.5 mL) was addeddiisopropyl azodicarboxylate (DIAD) (8 mg, 38 umol). The resultantyellow solution was stirred at room temperature for 4 h. After removalof the solvent, the crude material was purified by silica gelchromatography (hexane to 30% Hexane/EtOAc) to afford 13.4 mg (90%yield) of the desired product,6-chloro-4′-(2,6-difluoro-benzoylamino)-biphenyl-3-carboxylic acid2-methoxy-ethyl ester, as an off-white powder.

¹H-NMR (CDCl₃) δ ppm 3.40 (s, 3H), 3.70 (t, 2H, J=6), 4.45 (t, 2H, J=6),7.01 (t, 2H, J=8), 7.38-7.62 (m, 4H), 7.75 (d, 2H, J=8), 7.85 (s, 1H),7.95 (d, 1H, J=8), 8.05 (s, 1H); ESMS calcd for C₂₃H₁₈ClF₂NO₄: 445.2.found: 446.2 (M+H).

Compound 3:2,6-Difluoro-N-(2′-methyl-5′-oxazol-2-yl-biphenyl-4-yl)-benzamide

Compound 1 was hydrolyzed by heating it in a solution of LiOH to yield4′-(2,6-difluoro-benzoylamino)-6-methyl-biphenyl-3-carboxylic acid. Amixture of 4′-(2,6-difluoro-benzoylamino)-6-methyl-biphenyl-3-carboxylicacid (800 mg, 2.2 mmol), 2,2-diethoxy-ethylamine (0.32 mL, 2.2. mmol),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) (5 mmol) in dry DMF(5 mL) was stirred at room temperature for 24 h. The mixture was dilutedwith water (20 mL) and extracted with ethyl acetate (EtOAc) (2×20 mL).The organic extract was washed with water and dried. The oil obtained onconcentration of the organic layer was purified by flash chromatographedon silica gel to give4′-(2,6-difluoro-benzoylamino)-6-methyl-biphenyl-3-carboxylic acid(2,2-diethoxy-ethyl)-amide as colorless oil (0.68 g).

The oil of above was treated with a solution of P₂O₅ (1 g) in MeSO₃H (6mL) and was kept at 140° C. for 4 h. The mixture was poured onto ice,neutralized with Na₂CO₃ and extracted with EtOAc (2×50 mL). The oilobtained on concentration of the organic layer was purified by flashchromatographed on silica gel to give2,6-difluoro-N-(2′-methyl-5′-oxazol-2-yl-biphenyl-4-yl)-benzamide asyellowish solid (0.50 g).

¹H-NMR (DMSO-d₆) δ (ppm) 10.96 (br, 1H), 8.22 (s, 1H), 7.9 (m, 1H), 7.8(m, 3H), 7.6 (m, 1H), 7.4 (m, 4H), 7.2 (t, 2H, J=9), 2.32 (s, 3H); ESMSclcd for C₂₃H₁₆F₂N₂O₂: 390.1. Found: 391.1 (M+H)⁺.

Compound 4:4′-[(3,5-Difluoro-pyridine-4-carbonyl)-amino]-6-methyl-biphenyl-3-carboxylicacid methyl ester

A mixture of 4′-amino-6-methyl-biphenyl-3-carboxylic acid methyl ester(0.50 g), 3,5-difluoro-isonicotinic and EDC (0.80 g) in drydimethylformamide (DMF) (12 mL) was stirred at room temperature for 3 h.The mixture was diluted with water (40 mL) and extracted with EtOAc(2×50 mL). The oil obtained on concentration of the organic layer wasflash chromatographed on silica gel to give4′-[(3,5-difluoro-pyridine-4-carbonyl)amino]-6-methyl-biphenyl-3-carboxylicacid methyl ester (0.45 g) as white solid.

¹H-NMR (CDCl₃) δ (ppm) 8.6 (br, 1H), 8.41 (s, 2H), 7.9 (d, 2H, J=8), 7.7(d, 2H, J=8), 7.4 (m, 3H), 3.85 (s, 3H), 2.32 (s, 3H); ESMS clcd forC₂₁H₁₆F₂N₂O₃: 382.1. Found: 383.2 (M+H)⁺.

Compound 5:N-[4-(5-Chloro-2-methoxy-pyridin-3-yl)-phenyl]-2,6-difluoro-benzamide

Compound 5 was prepared by an analogous method as Compound 1 except that3-bromo-4-methyl-benzoic acid methyl ester was replaced with3-bromo-5-chloro-2-methoxypyridine.

¹H NMR (300 MHz, CDCl₃): 8.21 (s, 1H), 7.88-6.95 (m, 7H), 6.73 (s, 1H),3.93 (s, 3H). ESMS calcd (C₁₉H₁₃ClF₂N₂O₂):374.06. found: 375.1 (M+H).

Compound 6:2,6-Difluoro-N-[2′-methyl-5′-(1H-tetrazol-5-yl)-biphenyl-4-yl]-benzamide

N-(5′-cyano-2′-methyl-biphenyl-4yl)-2,6-difluoro-benzamide (Compound A)was prepared by an analogous method as Compound 1 except that3-bromo-4-methyl-benzoic acid methyl ester was replaced with2-bromo-4-cyano-toluene. A mixture of Compound A (348 mg 1 mmol), sodiumazide (78 mg 1.2 mmol), and ammonium chloride (65 mg, 1.2 mmol) in DMF(5 ml) was stirred and heated at 120° C. for 10 hr. After cooling thereaction mixture to room temperature, EtOAc (200 ml) was added, and themixture was washed with water (60 ml×3). The EtOAc layer wasconcentrated and the residue was subjected to silica gel chromatography(Hexanes:EtOAc, EtOAc:MeOH) to give the product,2,6-difluoro-N-[2′-methyl-5′-(1H-tetrazol-5yl)-biphenyl-4-yl]-benzamide(313 mg, yield 80%) as a off-white solid.

¹H NMR (300 MHz, CDCl₃): 7.96-6.97 (m, 10H), 2.37 (s, 3H). ESMS calcd(C₂₁H₁₅F₂N₅O):391.12. found: 392.1 (M+H).

Compound 10:3-Fluoro-N-(2′-methyl-5′-oxazol-2-yl-biphenyl-4-yl)-isonicotinamide

Bis(benzonitrile)dichloropalladium (0.03 mmol) and1,4-bis(diphenylphosphino)-butane (dppb, 0.03 mmol) in toluene (5 mL)were stirred under N₂ for 30 min. 2-(3-Iodo-4-methyl-phenyl)-oxazole (a,1.0 mmol) and3-fluoro-N-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-isonicotinamide(b, 1.0 mmol) was added followed by potassium carbonate solution (1M,1.0 mL) and ethanol (0.2 mL) and the mixture was heated at 90° C. for 12h. The mixture was loaded onto silica gel and purified by was flashchromatography to give3-fluoro-N-(2′-methyl-5′-oxazol-2-yl-biphenyl-4-yl)-isonicotinamide(Compound 10) as white solid (0.8 mmol).

¹H-NMR (CDCl₃) δ (ppm) 8.7 (m, 2H), 8.4 (br, 1H), 8.1 (t, 1H, J=6), 8.0(m, 2H), 7.7 (m, 3H), 7.4 (m, 2H), 7.2 (m, 2H), 2.34 (s, 3H); ESMS clcdfor C₂₂H₁₆FN₃O₂: 373.1. Found: 374.1 (M+H)⁺.

Compound 7:3-Methyl-N-(2′-methyl-5′-oxazol-2-yl-biphenyl-4-yl)-isonicotinamide,hydrochloride

¹H-NMR (DMSO-d₆) δ (ppm) δ 10.65 (br, 1H), 8.6 (m, 2H), 8.22 (s, 1H),7.8 (m, 4H), 7.4 (m, 5H), 2.39 (s, 3H), 2.32 (s, 3H); ESMS clcd forC₂₃H₂₀ClN₃O₂: 405.1. Found: 370.1 (M−Cl)⁺.

Compound 11:3,5-Difluoro-N-(2′-methyl-5′-oxazol-2-yl-biphenyl-4-yl)-isonicotin-amide

¹H-NMR (CDCl₃) δ (ppm) 9.47 (s, 2H), 8.2 (br, 1H), 7.9 (m, 2H), 7.7 (m,3H), 7.4 (m, 3H), 7.19 (s, 1H), 2.33 (s, 3H); ESMS clcd forC₂₂H₁₅F₂N₃O₂: 391.1. Found: 392.1 (M+H)⁺.

Compound 15:2,6-Difluoro-N-(2′-methyl-5′-thiazol-2-yl-biphenyl-4-yl)-benzamide

To a suspension of compound d (10 g) in benzene (700 ml), lawesson'sreagent (20 g) was added, the reaction was refluxed for 8 min in 100° C.oil bath. The mixture was filtered with silica gel funnel, and elutedwith CH₂Cl₂/EtOAc (1:1), and subjected to silical gel columnchromatography (5:1 Hexanes:Ethyl acetate) to give compound e (5.3 g).

To a solution of compound e (5.3 g) in THF (anhydrous) (50 ml) was addedbromoacetaldehyde diethyl acetal (10 ml), the mixture was refluxed for24 m hr. (Check by TLC). The solvents were evaporated and the residuewas purified with silica gel column chromatography to give f (2.8 g).

To a suspension of compound f (300 mg) toluene (100 ml) was addedcompound h (360 mg), dichlorobis(triphenylphosphine)palladium(II) (160mg), 1M Na₂CO₃ (900 μl), and ethanol (150 μl). The reaction was at 100°C. for 10 hours (check by TLC). The mixture was directly applied tosilica gel column chromatography to give compound 15 (315 mg).

¹H-NMR (CDCl₃) δ (ppm), 7.79-6.92 (m, 12H), 2.32 (s, 3H). ESMS clcd forC₂₃H₁₆F₂N₂OS: 406.10. Found: 407.1 (M+H)⁺.

Compound 14:3-Methyl-N-(2′-methyl-5′-thiazol-2-yl-biphenyl-4-yl)-isonicotinamide

To a solution of h (2 g) in methanol (100 ml) containing NH₃ (3 eq) wasbubbled with H₂S for 2 hr, after standing for another 10 hr, the solventwas evaporated to give crude j, which was used directly for the nextstep.

Alternatively, j can be prepared as follows. To a stirred suspension ofi (10 g) in benzene (700 ml) was added lawesson's reagent (20 g). Thereaction was refluxed for 8 min in 100° C. oil bath. The mixture wasfiltered with silica gel funnel, and eluted with CH₂Cl₂/EtOAc (1:1), andsubjected to silical gel column chromatography (5:1 Hexanes:Ethylacetate) to give j (5.3 g).

To a stirred solution of j (5.3 g) in THF (anhydrous, 50 ml) was addedbromoacetaldehyde diethyl acetal (10 ml). The mixture was refluxed andfollowed by TLC to determine when the reaction had gone to completion.After 24 hrs, the solvents were evaporated and the residue was purifiedwith silica gel column chromatography to give k (3.5 g).

To a stirred suspension of k (2.5 g) in toluene (500 ml) was added I(3.4 g), dichlorobis(triphenylphosphine)-palladium(II) (1.6 g), 1MNa₂CO₃ (7.5 ml), and ethanol (12.5 ml). The reaction mixture was stirredat 115° C. for 10 hours (check by TLC). After being cooled to roomtemperature, the mixture was directly applied to silica gel columnchromatography to give compound 14 (3.1 g).

¹H-NMR (CD₃Cl) δ (ppm), 8.53 (s, 1H), 8.49 (d, J=4.2, 1H), 8.11 (s, 1H),7.82-7.22 (m, 10H), 2.50 (s, 3H), 2.32 (s, 3H). ESMS clcd forC₂₃H₁₉N₃OS: 385.12. Found: 386.1. (M+H)⁺.

Compound 8:N-[4-(5-Chloro-2-methoxy-pyridin-3-yl)-phenyl]-2,6-difluoro-benzamide,hydrochloride

¹H-NMR (CD₃OD) δ (ppm), 8.25 (s, 1H), 8.84-7.13 (m, 7H), 7.01 (s, 1H),4.02 (s, 3H). ESMS clcd for C₁₉H₁₄Cl₂N₂O₂: 410.04. Found: 375.1(M+H—HCl)⁺.

Compound 17:3-Methyl-N-(2′-chloro-5′-oxazol-2-yl-biphenyl-4-yl)-isonicotin-amide,hydrochloride

¹H-NMR (CD₃OD) δ (ppm), 8.95 (s, 1H), 8.87-7.51 (m, 10H), 2.63 (s, 3H).ESMS clcd for C₂₂H₁₇Cl₂N₃O₂: 425.07. Found: 390.1 (M+H—HCl)⁺.

¹H-NMR (DMSOd₆) δ (ppm), 10.95 (s, 1H), 8.87-8.73 (m, 2H), 8.26 (s, 1H),8.01-7.42 (m, 8H), 7.41 (s, 1H), 2.50 (s, 3H).

Compound 16:3-Methyl-N-(2′-chloro-5′-oxazol-2-yl-biphenyl-4-yl)-isonicotinamide

¹H-NMR (DMSO-d₆) δ (ppm), 10.62 (s, 1H), 8.60-7.43 (m, 10H), 7.41 (s,1H), 2.39 (s, 1H). ESMS clcd for C₂₂H₁₆ClN₃O₂: 389.09. Found: 390.1(M+H)⁺.

¹H-NMR (CD₃Cl) δ (ppm), 10.67 (s, 1H), 8.58 (s, 1H), 8.46 (d, J=4.1,1H), 8.22 (s, 1H), 7.96-7.42 (m, 8H), 7.40 (s, 1H), 2.39 (s, 3H). ESMSclcd for C₂₂H₁₆ClN₃O₂: 389.09. Found: 390.1. (M+H)⁺.

Compound 19:3-Fluoro-N-[4-(5-chloro-2-methoxy-pyridin-3-yl)-phenyl]-isonicotinamide

¹H-NMR (CDCl₃) δ (ppm), 8.69-7.40 (m, 8H), 6.76 (s, 1H), 3.89 (s, 3H).ESMS clcd for C₁₈H₁₃ClFN₃O₂: 357.07. Found: 358.1 (M+H)⁺.

Compound 18:3-Methyl-N-[4-(5-chloro-2-methoxy-pyridin-3-yl)-phenyl]-isonicotinamide

¹H-NMR (CDCl₃) δ (ppm), 8.569-7.27 (m, 8H), 6.765 (s, 1H), 3.92 (s, 3H),2.22 (s, 3H). ESMS clcd for C₁₉H₁₆ClN₃O₂: 353.09. Found: 354.1 (M+H)⁺.

Compound 13:2,6-Difluoro-N-(2′-methoxy-5′-oxazol-2-yl-biphenyl-4-yl)-benzamide

¹H-NMR (CDCl₃) δ (ppm), 8.02 (s, 1H), 7.73-6.93 (m, 11H), 3.84 (s, 3H).ESMS clcd for C₂₃H₁₆F₂N₂O₃: 406.11. Found: 407.1 (M+H)⁺.

Compound 20:3-Methyl-N-[5′-(pyridin-3-yl)-Z-methyl-biphenyl-4-yl]-isonicotinamide

To a solution of 2-bromo-4-iodo-toluene (500 mg, 1.68 mmol),dichlorobis(triphenylphosphine)palladium (II) (Pd(PPh₃)₂Cl₂, 175 mg,0.25 mmol), and 3-pyridineboronic acid (200 mg, 1.62 mmol) in toluene (8mL) was added Na₂CO₃ (2 N, 1.0 mL) and ethanol (1.0 mL). The stirredmixture was heated up to 80° C. in the sealed tube for 24 hr. Thesolution was cooled to room temperature and diluted with H₂O (20 mL) andEtOAc (20 mL). The organic phase was dried over Na₂SO₄, concentrated,and chromatographed to give the pure product m (265 mg, 64%).

Suzuki Coupling Reaction:

To a solution of 3-(3-bromo-4-methylphenyl)-pyridine m (145 mg, 0.58mmol), dichlorobis(triphenylphosphine)palladium (II) (Pd(PPh₃)₂Cl₂, 60mg, 0.09 mmol), and 4-aminophenylboronic acid pinacol ester (130 mg,0.58 mmol) in toluene (4 mL) was added Na₂CO₃ (2 N, 0.3 mL) and ethanol(0.5 mL). The stirred mixture was heated up to 80° C. for 6 hr. Thesolution was cooled to room temperature and diluted with H₂O (10 mL) andEtOAc (10 mL). The organic phase was dried over Na₂SO₄, concentrated,and chromatographed to give n (90 mg, 60%).

Amide Coupling Reaction:

To a solution of 2′-methyl-5′-(pyridin-3-yl)biphenyl-4-amine n (40 mg,0.15 mmol) in DCM (3 mL) was added EDC (85 mg, 0.45 mmol) and3-methylisonicotinic acid (40 mg, 0.3 mmol). The solution was stirred atroom temperature for 6 hr before it was concentrated and chromatographedto give Compound 20 (50 mg, 88%).

¹H NMR (300 MHz, CDCl₃) δ 8.99 (s, 1H), 8.79-8.77 (m, 1H), 8.47-8.41 (m,3H), 7.90-7.86 (m, 1H), 7.75 (d, J=8.4 Hz, 2H), 7.49-7.26 (m, 7H), 2.47(s, 3H), 2.34 (s, 3H). ESMS calcd (C₂₅H₂₁N₃O): 379.1. found: 380.4(M+H).

Compound 21: 4-Methyl-[1,2,3]thiadazole-5-carboxylic acid[2′-methyl-5′-(pyridin-3-yl)-biphenyl-4-yl]-amide

¹H NMR (300 MHz, CDCl₃) δ 8.85 (d, J=2.4 Hz, 1H), 8.57-8.54 (m, 1H),7.91-7.87 (m, 2H), 7.68-7.65 (m, 2H), 7.52-7.34 (m, 6H), 3.00 (s, 3H),2.33 (s, 3H). ESMS calcd (C₂₂H₁₈N₄OS): 386.1. found: 387.2 (M+H).

Compound 22:2,6-Difluoro-N-[2′-methyl-5′-(pyridine-3-yl)-biphenyl-4-yl]-benzamide

Compound 22 was prepared by a method analogous to that described for theamide coupling reaction of Compound 20.

¹H NMR (300 MHz, CDCl₃) δ 8.81 (s, 1H), 8.55-8.50 (m, 1H), 8.24 (s, 1H),7.91-7.85 (m, 1H), 7.76-7.71 (m, 2H), 7.51-7.31 (m, 7H), 7.02-6.93 (m,2H), 2.32 (s, 3H); ESMS calcd (C₂₅H₁₈F₂N₂O): 400.1. found: 401.1 (M+H).

Compound 23:2,6-Difluoro-N-[2′-methyl-5′-(pyridine-2-yl)-biphenyl-4-yl]-benzamide

¹H NMR (300 MHz, CDCl₃) δ 8.68-8.62 (m, 1H), 8.11 (s, 1H), 7.82-7.64 (m,6H), 7.41-7.16 (m, 5H), 6.99-6.86 (m, 2H), 2.33 (s, 3H); ESMS calcd(C₂₅H₁₈F₂N₂O): 400.1. found: 401.0 (M+H).

Compound 24: 4-Methyl-[1,2,3]thiadazole-5-carboxylic acid[2′-methyl-5′-(pyridin-2-yl)-biphenyl-4-yl]-amide

¹H NMR (300 MHz, CDCl₃) δ 8.67-8.64 (m, 1H), 8.17 (s, 1H), 7.84-7.63 (m,5H), 7.39-7.36 (m, 3H), 7.26-7.20 (m, 1H), 2.94 (s, 3H), 2.33 (s, 3H);ESMS calcd (C₂₂H₁₈N₄OS): 386.1. found: 387.2 (M+H).

Compound 30:2,6-Difluoro-N-[2′-methyl-5-(pyridine-4-yl)-biphenyl-4-yl]-benzamide

¹H NMR (300 MHz, CDCl₃) δ 8.63-8.58 (m, 2H), 8.13 (s, 1H), 7.77-7.36 (m,10H), 7.04-6.96 (m, 2H), 2.33 (s, 3H); ESMS calcd (C₂₅H₁₈F₂N₂O): 400.1.found: 401.1 (M+H).

Compound 25: 4-Methyl-[1,2,3]thiadazole-5-carboxylic acid[2′-methoxy-5′-(oxazol-5-yl)-biphenyl-4-yl]-amide

The mixed solution of 3-bromo-4-methoxybenzaldehyde (200 mg, 0.93 mmol)in methanol (4 mL) was added tosylmethyl isocyanide (200 mg, 1.02 mmol)and K₂CO₃ (260 mg, 1.88 mmol). The reaction was stirred at roomtemperature for 5 min before heated to 80° C. in the sealed tube. After30 min, the solution was cooled to room temperature and concentrated.Column chromatography afforded 5-(3-bromo-4-methoxyphenyl)oxazole (o)(190 mg, 80%). Following the procedures analogous to the Suzuki andamide coupling reactions described for Compound 20, Compound 25 wasprepared.

¹H NMR (300 MHz, CD₃OD) δ 8.22 (s, 1H), 7.74-7.65 (m, 4H), 7.57-7.54 (m,2H), 7.43 (s, 1H), 7.20-7.16 (m, 1H), 3.87 (s, 3H), 2.89 (s, 3H); ESMScalcd (C₂₀H₁₆N₄O₃S): 392.1. found: 393.1 (M+H).

Compound 27:2,6-Difluoro-N-[2′-methyl-5′-(oxazol-5-yl)-biphenyl-4-yl]-benzamide

¹H NMR (300 MHz, CD₃OD) δ 8.19 (s, 1H), 7.73-7.40 (m, 8H), 7.16-7.09 (m,3H), 3.84 (s, 3H); ESMS calcd (C₂₃H₁₆F₂N₂O₃): 406.1. found: 407.0 (M+H).

Compound 26:3-Methyl-N-[5′-(oxazol-5-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide

¹H NMR (300 MHz, CDCl₃) δ 8.57-8.54 (m, 2H), 7.89 (s, 1H), 7.81 (s, 1H),7.73-7.70 (m, 2H), 7.57-7.51 (m, 2H), 7.40-7.31 (m, 5H), 2.52 (s, 3H),2.31 (s, 3H); ESMS calcd (C₂₃H₁₉N₃O₂): 369.1. found: 370.2 (M+H).

Compound 29:3-Methyl-N-[5′-(oxazol-5-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide,hydrochloride salt

¹H NMR (300 MHz, (CD₃)₂SO) δ 10.97 (s, 1H), 8.86 (s, 1H), 8.82 (d, J=5.4Hz, 1H), 8.42 (s, 1H), 7.96 (d, J=5.4 Hz, 1H), 7.82 (d, J=8.7 Hz, 2H),7.69 (s, 1H), 7.63-7.54 (m, 2H), 7.43-7.38 (m, 2H), 4.10 (brs, 1H), 3.31(s, 3H), 2.47 (s, 3H); ESMS calcd (C₂₃H₁₉N₃O₂): 369.1. found: 370.1(M+H).

Compound 35:2,6-Difluoro-N-[2′-methyl-5′-(3-methyl-isoxazole-5-yl)-biphenyl-4-yl]-benzamide

A solution of 3′-bromo-4′-methylacetophenone (1 g, 4.69 g) inN,N-dimethylacetamide dimethyl acetal (2.5 mL) was refluxed at 100° C.for 12 hr. The solvent was removed and the residue and hydroxylaminehydrochloride (490 mg, 7.1 mmol) was dissolved in ethanol (10 mL). Thesolution was refluxed at 90° C. for 2 hr before it was concentrated.Column chromatography afforded compound q in 65% overall yield. Compound35 was obtained following a Suzuki coupling and amide couplingprocedures analogous to that described for compound 20.

¹H NMR (300 MHz, CDCl₃) δ 7.82-7.61 (m, 5H), 7.49-7.36 (m, 4H),7.04-6.96 (m, 2H), 6.32 (s, 1H), 2.37 (s, 3H), 2.34 (s, 3H); ESMS calcd(C₂₄H₁₈F₂N₂O₂): 404.1. found: 405.1 (M+H).

Compound 37:2,6-Difluoro-N-[2′-methyl-5′-(3-methyl-1H-pyrazol-5-yl)-biphenyl-4-yl]-benzamide

A solution of 3′-bromo-4′-methylacetophenone (1 g, 4.69 g) inN,N-dimethylacetamide dimethyl acetal (2.5 mL) was refluxed at 100° C.for 12 hr. The solvent was removed and the residue and hydrazinemonohydrate (355 mg, 7.1 mmol) was dissolved in ethanol (10 mL). Thesolution was refluxed at 90° C. for 1 hr before it was concentrated.Column chromatography afforded compound r in 75% overall yield. Compound37 was obtained by a Suzuki coupling reaction analogous to thatdescribed for Compound 20.

¹H NMR (300 MHz, (CD₃)₂SO) δ 10.91 (s, 1H), 7.79-7.21 (m, 11H), 6.41 (s,1H), 2.22 (s, 6H); ESMS calcd (C₂₄H₁₉F₂N₃O): 403.1. found: 404.1 (M+H).

Compound 36:3-Methyl-N-[5′-(3-methyl-isoxazol-5-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide

¹H NMR (300 MHz, CD₃OD) δ 8.54-8.51 (m, 2H), 7.81-7.28 (m, 8H), 6.62 (s,1H), 2.48 (s, 3H), 2.32 (s, 6H); ESMS calcd (C₂₄H₂₁N₃O₂): 383.1. found:484.2 (M+H).

Compound 32: 3-Methyl-N-[5′-(isoxazol-5-yl)-2′-methyl-biphenyl-4-yl]isonicotinamide

¹H NMR (300 MHz, CDCl₃) δ 8.59-8.56 (m, 2H), 8.27 (d, J=1.8 Hz, 1H),7.73-7.65 (m, 5H), 7.40-7.37 (m, 4H), 6.49 (d, J=1.8 Hz, 1H), 2.52 (s,3H), 2.34 (s, 3H); ESMS calcd (C₂₃H₁₉N₃O₂): 369.1. found: 370.2 (M+H).

Compound 34:3-Methyl-N-[5′-(isoxazol-5-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide,HCl salt

¹H NMR (300 MHz, (CD₃)₂SO) δ 10.84 (s, 1H), 8.78 (s, 1H), 8.75 (d, J=5.1Hz, 1H), 8.64-8.63 (m, 1H), 7.83-7.69 (m, 5H), 7.48-7.41 (m, 3H), 7.04(d, J=1.5 Hz, 1H), 3.72 (brs, 1H), 2.45 (s, 3H), 2.30 (s, 3H);); ESMScalcd (C₂₃H₁₉N₃O₂): 369.1. found: 370.1 (M+H).

Compound 33: 4-Methyl-[1,2,3]thiadazole-5-carboxylic acid[2′-methyl-5′-(isoxazol-5-yl)-biphenyl-4-yl]-amide

¹H NMR (300 MHz, CDCl₃) δ 8.39 (s, 1H), 8.24 (d, J=1.8 Hz, 1H),7.71-7.63 (m, 4H), 7.38-7.34 (m, 3H), 6.50 (d, J=2.1 Hz, 1H), 2.95 (s,3H), 2.31 (s, 3H); ESMS calcd (C₂₀H₁₆N₄O₂S): 376.1. found: 377.1 (M+H).

Compound 38:2,6-Difluoro-N-[2′-methyl-5′-(4-methyl-thiazol-2-yl)-biphenyl-4-yl]-benzamide

A solution of 3-bromo-4-methyl-benzonitrile (500 mg, 2.55 mmol) inammonia solution (2 M in ethanol, 10 mL) was bubbled with H₂S gas slowlyfor 1 hr. The solution was stirred for 3 hr at room temperature beforethe nitrogen was bubbled through the solution to remove H₂S. Thesolution was concentrated to give the crude s (450 mg) which was useddirectly in the next reaction.

The solution of s (100 mg, 0.43 mmol) and 1-chloropropan-2-one (200 μL,2.5 mmol) in ethanol (2 mL) was refluxed at 85° C. for 10 hr. Thesolvent was removed and column chromatography afforded t (60 mg, 52%).Compound 38 was obtained by a Suzuki coupling reaction analogous to thatdescribed for Compound 20.

¹H NMR (300 MHz, CDCl₃) δ 7.93 (s, 1H), 7.80-7.68 (m, 4H), 7.40-7.29 (m,4H), 7.03-6.96 (m, 2H), 6.82 (s, 1H), 2.48 (s, 3H), 2.31 (s, 3H); ESMScalcd (C₂₄H₁₈F₂N₂OS): 420.1. found: 421.1 (M+H).

Compound 39:2,6-Difluoro-N-[2′-methyl-5′-(4-trifluoromethyl-thiazol-2-yl)-biphenyl-4-yl]-benzamide

A solution of s (100 mg, 0.43 mmol) and3-bromo-1,1,1-trifluoropropan-2-one (270 μL, 2.57 mmol) in ethanol (4mL) was refluxed at 85° C. for 4 hr. The solvent was removed and theresidue was dissolved in dichloromethane (4 mL) with TEA (120 μL, 0.86mmol) and TFAA (120 μL, 0.86 mmol). The reaction was stirred at roomtemperature for 30 min before the solution was concentrated. Columnchromatography afforded t (100 mg, 68%). Compound 39 was obtained by aSuzuki coupling reaction analogous to that described for Compound 20.

¹H NMR (300 MHz, CDCl₃) δ 7.88-7.64 (m, 6H), 7.48-7.38 (m, 4H),7.07-7.02 (m, 2H), 2.33 (s, 3H); ESMS calcd (C₂₄H₁₅F₅N₂OS): 474.1.found: 475.0 (M+H).

Compound 40:3-Methyl-N-[5′-(4-methyl-thiazol-2-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide

¹H NMR (300 MHz, CD₃OD) δ 8.54-8.51 (m, 2H), 7.81-7.76 (m, 2H),7.68-7.52 (m, 5H), 7.41-7.38 (m, 2H), 7.11 (s, 1H), 2.47 (s, 3H), 2.45(s, 3H), 2.34 (s, 3H); ESMS calcd (C₂₄H₂₁N₃OS): 399.1. found: 400.1(M+H).

Compound 42:2,6-Difluoro-N-[2′-methyl-5′-(5-oxo-4,5-dihydro-[1,2,4]-oxadiazol-3-yl)-biphenyl-4-yl]-benzamide

A mixed solution of 3-bromo-4-methylbenzonitrile (500 mg, 2.6 mmol) andNH₂OH (50% in H₂O, 0.4 mL, 6.5 mmol) in EtOH (3 mL) was refluxed in asealed tube at 85° C. for 5 hr. The solvent was removed and the residuewas dissolved in THF (4 mL). To the solution was added pyridine (0.31mL, 3.8 mmol) and 2-ethylhexy chloroformate (0.75 mL, 3.8 mmol) at 0° C.and stirred at this temperature for 1 hr. The organic phase was washedwith H₂O and brine and concentrated to give a residue that was dissolvedin xylene (5 mL). The solution was refluxed at 110° C. for 12 hr,concentrated, and partitioned between water and ethyl acetate. Theorganic phase was separated, washed with water and brine, and dried.Concentration followed by column chromatography afforded u (350 mg).

Following a Suzuki coupling procedure analogous to that described forCompound 20, Compound 42 was obtained as solid.

¹H NMR (300 MHz, (CD₃)₂SO) δ 10.92 (s, 1H), 7.82-7.19 (m, 11H), 2.24 (s,3H); ESMS calcd (C₂₂H₁₅F₂N₃O₃): 407.1. found: 408.1 (M+H).

Compound 9:3-Methyl-N-(2′-methyl-5′-thiazol-2-yl-biphenyl-4-yl)-isonicotinamide,hydrochloride salt

To a stirred suspension of Compound 42 (3 g) in methanol (60 mL) wasadded HCl (2 eq) in methanol (40 mL). Ether (200 mL) was then added tothe resultant solution at room temperature. After 2 hrs, precipitateswere collected and dried to give the title compound as a solid (3.1 g).

¹H-NMR (DMSOd₆) δ (ppm), 11.12 (s, 1H), 8.99-7.37 (m, 12H), 7.51-7.38(m, 3H), 2.51 (s, 3H), 2.27 (s, 3H). ESMS clcd for C₂₃H₂₀ClN₃OS: 421.10.Found: 386.1 (M−HCl+H)⁺.

General Method for the Synthesis of Compounds 41, 43, 47, and 52:

To a solution of 3-iodo-4-methylaniline (1 g, 4.29 mmol) in H₂O (25 mL)was added H₂SO₄ (0.5 M, 25 mL). The solution was heated to 80° C. untilall solid dissolved. Then the reaction was cooled to 0° C. and NaNO₂(444 mg, 6.39 mmol) was added in small portions. After 2 hr at thistemperature, urea (126 mg, 2.1 mmol) was added at 0° C. The solution wasallowed to warm up to room temperature and H₂SO₄ (0.5 M, 25 mL) wasadded. The reaction was refluxed for 30 min and cooled down to roomtemperature. The solution was extracted with EtOAc and Et₂O and thecombined organic phases were dried over Na₂SO₄, concentrated, andchromatographed to give the pure product v (800 mg, 80%).

2,6-Difluoro-N-(5′-hydroxy-2′-methylbiphenyl-4-yl)benzamide), w, wasprepared from v and2,6-Difluoro-N-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-benzamidefollowing Suzuki coupling procedure described above for Compound 20.

¹H NMR (300 MHz, CDCl₃) δ 7.83 (s, 1H), 7.68-7.62 (m, 2H), 7.42-7.28 (m,3H), 7.11-6.85 (m, 3H), 6.78-6.72 (m, 2H), 2.18 (s, 3H); ESMS calcd(C₂₀H₁₅F₂NO₂): 339.1. found: 340.1 (M+H).

To a solution of w (1 g, 2.94 mmol) and pyridine (355 μL, 4.39 mmol) indichloromethane (DCM) (15 mL) was added trifluoromethanesulfonicanhydride (545 μL, 3.24 mmol) in drop wise at 0° C. After 10 min at thistemperature, the solvent was removed and column chromatography afforded(4′-(2,6-difluorobenzamido)-6-methylbiphenyl-3-yltrifluoromethanesulfonate, x, (1.17 g, 85%).

¹H NMR (300 MHz, CDCl₃) δ 7.78-7.66 (m, 3H), 7.49-7.31 (m, 4H),7.20-7.11 (m, 2H), 7.08-6.99 (m, 2H), 2.28 (s, 3H); ESMS calcd(C₂₁H₁₄F₅NO₄S): 471.1. found: 472.0 (M+H).

Compound 41, Compound 47 and Compound 52 were synthesized from x using aSuzuki coupling analogous to that described for Compound 20. Compound 43was prepared by nucleophilic substitution of the aromatic triflate bymorpholino:

Compound 41:2,6-Difluoro-N-[2′-methyl-5′-(1H-pyrrol-2-yl)-biphenyl-4-yl]-benzamide

Compound 41 was prepared from w following a Suzuki coupling reactionanalogous to that described for Compound 20.

¹H NMR (300 MHz, CDCl₃) δ 8.61 (s, 1H), 7.93 (s, 1H), 7.67-7.64 (m, 2H),7.41-7.22 (m, 5H), 7.02-6.96 (m, 2H), 6.82-6.80 (m, 1H), 6.51-6.49 (m,1H), 6.30-6.28 (m, 1H), 2.25 (s, 3H); ESMS calcd (C₂₄H₁₈F₂N₂O): 388.1.found: 389.1 (M+H).

Compound 43:2,6-Difluoro-N-[2′-methyl-5′-(morpholino-4-yl)-biphenyl-4-yl]-benzamide

¹H NMR (300 MHz, CDCl₃) δ 8.05 (s, 1H), 7.70-7.65 (m, 2H), 7.43-7.15 (m,4H), 7.02-6.95 (m, 2H), 6.85-6.78 (m, 2H), 3.85 (t, J=5.1 Hz, 4H), 3.13(t, J=5.1 Hz, 4H), 2.19 (s, 3H); ESMS calcd (C₂₄H₂₂F₂N₂O₂): 408.2.found: 409.3 (M+H).

Compound 47:3-Methyl-N-[2′-methoxy-5′-(furan-2-yl)-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CD₃Cl) δ (ppm), 8.52 (s, 1H), 8.49 (d, J=4.2, 1H), 7.95-7.28 (m,11H), 3.79 (s, 3H), 2.46 (s, 3H). ESMS clcd for C₂₄H₂₀N₂O₃: 384.15.Found: 385.2. (M+H)⁺.

Compound 52:3-Methyl-N-[5′-(thien-2-yl)-2′-methoxy-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CD₃Cl) δ (ppm), 8.53 (s, 1H), 8.49 (d, J=4.1, 1H), 7.75-7.35 (m,11H), 3.79 (s, 3H), 2.47 (s, 3H). ESMS clcd for C₂₄H₂₀N₂O₂S: 400.12.Found: 401.1. (M+H)⁺.

General Method for the Synthesis of Compounds 46, 68, 69, 70, and 71:

3-Iodo-4-methyl benzoic acid methyl ester was treated with hydrazine toform 3-Iodo-4-methyl benzoic acid hydrazide.2-(3-Iodo-4-methyl-phenyl)-[1,3,4]oxadiazole was prepared from3-iodo-4-methyl benzoic acid hydrazide according to a method analogousto that described in J. of Medicinal Chemistry (2001), 44(8):1268-85,the entire teachings of which are incorporated herein by reference.Compounds 46, 68, 69, 70, and 71 were prepared via an amide couplingreaction analogous to that described in step A of the synthesis ofCompound 1, followed by a Suzuki coupling reaction analogous to thatdescribed in step B of the synthesis of Compound 1. The amide couplingreaction and Suzuki coupling reaction are shown for Compound 69 inScheme XX below:

Compound 46:3-Methyl-N-[2′-methyl-5′-([1,3,4]oxadiazol-2-yl)-biphenyl-4-yl]-isonicotinamide,HCl salt

¹H-NMR (CD₃OD) δ (ppm), 9.02 (s, 1H), 8.89 (s, 1H), 8.61 (d, J=5.2, 1H),8.18 (d, J=5.2, 1H), 7.91 (m, 2H), 7.82 (d, J=7.6, 2H), 7.53 (d, J=6.4,1H), 7.42 (d, J=7.6, 2H), 2.63 (s, 3H), 2.38 (s, 3H). ESMS clcd forC₂₂H₁₉ClN₄O₂: 406.12. Found: 371.1 (M−HCl+H)⁺.

Compound 68:3,5-Difluoro-N-[2′-methyl-5′-([1,3,4]oxadiazol-2-yl)-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CDCl₃) δ, 8.52 (s, 2H), 8.43 (s, 1H), 7.99-7.92 (m, 2H), 7.83(s, 1H), 7.71 (d, J=7.5, 2H), 7.43 (m, 1H), 7.38 (d, J=7.5, 2H), 2.38(s, 3H). ESMS clcd for C₂₁H₁₄F₂N₄O₂: 392.11. Found: 393.1. (M+H)⁺.

Compound 69:2,6-Difluoro-N-[2′-methyl-5′-([1,3,4]oxadiazol-2-yl)-biphenyl-4-yl]-benzamide

¹H-NMR (CDCl₃) δ (ppm), 8.42 (s, 1H), 8.02-7.34 (m, 8H), 7.11 (m, 2H),2.36 (s, 3H). ESMS clcd for C₂₂H₁₅F₂N₃O₂: 391.11. Found: 392.1 (M+H)⁺.

Compound 70:3-Methyl-N-[2′-methyl-5′-([1,3,4]oxadiazol-2-yl)-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CD₃Cl) δ (ppm), 8.60-8.52 (m, 2H), 8.42 (s, 1H), 7.99 (m, 3H),7.71 (d, J=7.6, 2H), 7.44-7.32 (m, 3H), 7.91 (m, 2H), 2.52 (s, 3H), 2.36(s, 3H). ESMS clcd for C₂₂H₁₈N₄O₂: 370.14. Found: 371.1 (M+H)⁺.

Compound 71:3-Fluoro-N-[2′-methyl-5′-([1,3,4]oxadiazol-2-yl)-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CDCl₃) δ, 8.72-7.84 (m, 7H), 7.73 (d, J=7.5, 2H), 7.42 (m, 1H),7.38 (d, J=7.5, 2H), 2.38 (s, 3H). ESMS clcd for C₂₁H₁₅FN₄O₂: 374.12.Found: 375.1. (M+H)⁺.

General Method for the Synthesis of Compounds 53, 59, 60, and 61:

A solution of 2-bromo-4-cyano-toluene and hydrazine was treated withhydrogen sulfide to form 2-3-bromo-4-methyl-thiobenzoic acid hydrazide.2-(3-Bromo-4-methyl-phenyl)-[1,3,4]thiadiazole was prepared from2-3-bromo-4-methyl-thiobenzoic acid hydrazide according to a methodanalogous to that described in J. of the American Chemical Society(1955), 77:1148, the entire teachings of which are incorporated hereinby reference. Compounds 53, 59, 60, and 61 were prepared via an amidecoupling reaction analogous to that described in step A of the synthesisof Compound 1, followed by a Suzuki coupling reaction (see step B of thesynthesis of Compound 1) analogous to that as shown for Compound 69 inScheme XX above.

Compound 53:3,5-Difluoro-N-[5′-([1,3,4]thiadiazol-2-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CDCl₃1) δ, 9.12 (s, 1H), 8.53 (s, 2H), 7.93-7.79 (m, 3H), 7.65(d, J=7.5, 2H), 7.38 (d, J=7.5, 2H), 2.37 (s, 3H). ESMS clcd forC₂₁H₁₄F₂N₄OS: 408.09. Found: 409.1. (M+H)⁺.

Compound 59:2,6-Difluoro-N-[2′-methyl-5′-([1,3,4]thiadiazol-2-yl)-biphenyl-4-yl]-benzamide

¹H-NMR (CD₃Cl) δ (ppm), 9.14 (s, 1H), 7.96-7.15 (m, 8H), 7.04 (m, 2H),2.37 (s, 3H). ESMS clcd for C₂₂H₁₅ClF₂N₃OS: 407.09. Found: 408.1.(M+H)⁺.

Compound 60:3-Methyl-N-[5′-([1,3,4]thiadiazol-2-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CD₃Cl) δ (ppm), 9.05 (s, 1H), 8.49 (m, 2H), 8.24 (s, 1H),7.94-7.32 (m, 8H), 2.48 (s, 3H), 2.39 (s, 3H). ESMS clcd for C₂₂H₁₈N₄OS:386.12. Found: 387.1. (M+H)⁺.

Compound 61:3-Fluoro-N-[5′-([1,3,4]thiadiazol-2-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CDCl₃) δ (ppm), 9.81 (s, 1H), 8.72-7.86 (m, 6H), 7.74 (d, J=7.5,2H), 7.41 (d, J=7.5, 2H), 2.37 (s, 3H). ESMS clcd for C₂₁H₁₅FN₄OS:390.10. Found: 391.1 (M+H)⁺.

General Method for the Synthesis of Compounds 62, 63, and 64:

2-(3-Bromo-4-methyl-phenyl)-5-amino [1,3,4]thiadiazole was preparedaccording to a method analogous to that described in Suzuki, et al.,Chem. Pharm. Bull. (1992), 40:357-363, the entire teachings of which areincorporated herein by reference. Compounds 62, 63, and 64 were preparedvia an amide coupling reaction analogous to that described in step A ofthe synthesis of Compound 1, followed by a Suzuki coupling reaction (seestep B of the synthesis of Compound 1) analogous to that as shown forCompound 69 in Scheme XX above.

# Chemical Name Structure ESMS 62 2,6-Difluoro-N-[2′-methyl-5′-(5-amino-[1,3,4]thiadiazol-2-yl)-biphenyl-4-yl]- benzamide

422.10 63 2,6-Difluoro-N-{2′-methyl-5′-[5-(N,N-dimethylamino)-[1,3,4]thiadiazol-2-yl]-biphenyl-4-yl}-N-methyl-benzamide

464.15 64 2,6-Difluoro-N-{2′-methyl-5′-[5-(N,N-dimethylamino)-[1,3,4]thiadiazol- 2-yl]-biphenyl-4-yl}-benzamide

450.13

General Method for the Synthesis of Compounds 66 and 67:

3-(3-Bromo-4-methyl-phenyl)-[1,2,4]triazole was prepared according to amethod analogous to that described in Organic Letters (2004),6(7):1111-1114; J. of Chemistry (2002), 67(10):3266-3271; EuropeanPatent Application No. 636625, or J. Heterocylic Chem. (1988),25(4):1151-1154, the entire teachings of each of thes references areincorporated herein by reference. Compounds 66 and 67 were prepared viaan amide coupling reaction analogous to that described in step A of thesynthesis of Compound 1, followed by a Suzuki coupling reaction (seestep B of the synthesis of Compound 1) analogous to that as shown forCompound 69 in Scheme XX above.

General Method for the Synthesis of Compounds 72, 73, 74, and 75:

3-Bromo-4-methyl-benzoic acid hydrazide was prepared by treating3-bromo-4-methyl-benzoic acid methyl ester with hydrazine and heat.3-Bromo-4-methyl-benzoic acid hydrazide was then prepared by heating itwith isothiocyanate in ethanol to form intermediate y. Intermediate ywas cyclized to form5-(3-bromo-4-methyl-phenyl)-4-methyl-4H-[1,2,4]triazole-3-thiol byheating it in an aqueous solution containing 2 molar equivalents ofNaOH. The mercapto group was then methylated by treating5-(3-bromo-4-methyl-phenyl)-4-methyl-4H-[1,2,4]-triazole-3-thiol withCH₂N₂ to form5-(3-bromo-4-methyl-phenyl)-4-methyl-5-methylsulfanyl-4H-[1,2,4]triazole.Compounds 72, 73, 74, and 75 were prepared via an amide couplingreaction analogous to that described in step A of the synthesis ofCompound 1, followed by a Suzuki coupling reaction (see step B of thesynthesis of Compound 1) analogous to that as shown for Compound 69 inScheme XX above.

# Chemical Name Structure ESMS 72 2,6-Difluoro-N-[2′-methyl-5′-(4-methyl-5-methylsulfanyl- 4H-[1,2,4]triazol-3-yl)-biphenyl-4-yl]-benzamide

450.13 75 3,5-Difluoro-N-[2′-methyl- 5′-(4-methyl-5-methylsulfanyl-4H-[1,2,4]triazol-3-yl)-biphenyl- 4-yl]-isonicotinamide

451.13 73 3-Fluoro-N-[2′-methyl- 5′-(4-methyl-5-methylsulfanyl-4H-[1,2,4]triazol-3-yl)-biphenyl- 4-yl]-isonicotinamide

433.14 74 3-Methyl-N-[2′-methyl- 5′-(4-methyl-5-methylsulfanyl-4H-[1,2,4]triazol-3-yl)-biphenyl- 4-yl]-isonicotinamide

429.16

The following examples were prepared using procedures analogous to thosedescribed above:

Compound 76: 4-Methyl-[1,2,3]thiadazole-5-carboxylic acid[2′-methyl-5′-(thiazol-2-yl)-biphenyl-4-yl]-amide

¹H-NMR (CD₃OD) δ (ppm), 7.88-7.72 (m, 5H), 7.59 (d, J=4.8, 1H),7.51-7.38 (m, 3H), 2.84 (s, 3H), 2.31 (s, 3H). ESMS clcd forC₂₀H₁₆N₄OS₂: 392.08. Found: 393.1 (M+H)⁺.

Compound 77:2,6-Difluoro-N-[2′-methyl-5′-(oxazol-4-yl)-biphenyl-4-yl]-benzamide

¹H-NMR (CD₃OD) δ (ppm), 7.88-7.72 (m, 5H), 7.59 (d, J=4.8, 1H),7.51-7.38 (m, 3H), 2.84 (s, 3H), 2.31 (s, 3H). ESMS clcd forC₂₃H₁₆F₂N₂O₂: 390.12. Found: 391.1 (M+H)⁺.

Compound 56:2,6-Difluoro-N-[2′-chloro-5′-(thiazol-2-yl)-biphenyl-4-yl]-benzamide

¹H-NMR (CD₃Cl) δ (ppm), 7.99-7.32 (m, 10H), 7.11 (m, 2H). ESMS clcd forC₂₂H₁₃ClF₂N₂OS: 426.04. Found: 427.1. (M+H)⁺.

Compound 55:3-Methyl-N-[5′-(thiazol-2-yl)-2′-chloro-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CD₃Cl) δ (ppm), 8.51 (s, 1H), 8.48 (d, J=4.2, 1H), 8.41 (s, 1H),7.95-7.36 (m, 10H), 2.51 (s, 3H). ESMS clcd for C₂₂H₁₆ClN₃OS: 405.07.Found: 406.1. (M+H)⁺.

Compound 50:3-Methyl-N-[5′-(oxazol-2-yl)-2′-methoxy-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CD₃Cl) δ (ppm), 8.56 (s, 1H), 8.02 (s, 1H), 7.75-7.03 (m, 10H),3.86 (s, 3H), 2.53 (s, 3H). ESMS clcd for C₂₃H₁₉N₃O₃: 385.14. Found:386.2. (M+H)⁺.

Compound 31:2,6-Difluoro-N-[2′-chloro-5′-(oxazol-2-yl)-biphenyl-4-yl]-benzamide

¹H-NMR (CD₃Cl) δ (ppm), 7.98-7.32 (m, 10H), 7.03 (m, 2H). ESMS clcd forC₂₂H₁₃ClF₂N₂O₂: 410.06. Found: 411.1. (M+H)⁺.

Compound 45:3,5-Difluoro-N-[5′-(oxazol-2-yl)-2′-chloro-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CDCl₃) δ, 8.53 (s, 2H), 8.11-7.28 (m, 9H). ESMS clcd forC₂₁H₁₂ClF₂N₃O₂: 411.06. Found: 412.1. (M+H)⁺.

Compound 48:3-Methyl-N-[5′-(oxazol-2-yl)-2′-(N,N-dimethylamino)-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CDCl₃) δ, 7.75-7.40 (m, 12H), 8.53 (s, 2H), 7.93-7.79 (m, 3H),2.43 (s, 3H), 1.36 (s, 6H). ESMS clcd for C₂₄H₂₂N₄O₂: 398.17. Found:399.1. (M+H)⁺.

Compound 51:3,5-Difluoro-N-[5′-(oxazol-2-yl)-2′-methoxy-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CDCl₃) δ (ppm), 8.40 (s, 2H), 7.81-7.30 (m, 9H), 3.84 (s, 3H).ESMS clcd for C₂₁H₁₄F₂N₁₄OS: 407.11. Found: 408.1. (M+H)⁺.

Compound 54:3-Fluoro-N-[5′-(thiazol-2-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CDCl₃) δ, 9.12 (s, 1H), 8.53 (s, 2H), 7.93-7.79 (m, 3H), 7.65(d, J=7.5, 2H), 7.38 (d, J=7.5, 2H), 2.37 (s, 3H). ESMS clcd forC₂₂H₁₆FN₃OS: 389.10. Found: 390.1. (M+H)⁺.

Compound 57:3,5-Difluoro-N-[5′-(thiazol-2-yl)-2′-chloro-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CDCl₃) δ, 8.45 (s, 2H), 7.95-7.41 (m, 9H). ESMS clcd forC₂₁H₁₂ClF₂N₃O₂: 427.04. Found: 428.1. (M+H)⁺.

Compound 58:3-Fluoro-N-[5′-(thiazol-2-yl)-2′-chloro-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CD₃OD) δ, 8.62 (s, 1H), 8.53 (d, J=4.1, 1H), 7.99-7.42 (m, 10H).ESMS clcd for C₂₁H₁₃ClFN₃OS: 408.09. Found: 409.1. (M+H)⁺.

Compound 78:3-Fluoro-N-[2′-methyl-5′-(oxazol-4-yl)-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CDCl₃) δ, 8.73-7.27 (m, 12H), 2.38 (s, 3H). ESMS clcd forC₂₂H₁₆FN₃O₂: 373.12. Found: 374.1. (M+H)⁺.

Compound 79:3-Methyl-N-[2′-methyl-5′-(oxazol-4-yl)-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CDCl₃) δ, 8.63 (s, 1H), 7.92 (s, 1H), 7.85-7.28 (m, 10H), 2.53(s, 3H), 2.36 (s, 3H). ESMS clcd for C₂₃H₁₉N₃O₂: 369.15. Found: 370.1.(M+H)⁺.

Compound 81:2,6-Difluoro-N-[2′-methyl-5′-(1H-tetrazol-5-yl)-biphenyl-4-yl]-benzamide,hydrochloride salt

¹H-NMR (CDCl₃) δ, 7.95-7.24 (m, 8H), 7.11-7.01 (m, 2H), 2.38 (s, 3H).ESMS clcd for C₂₁H₁₆ClF₂N₅O: 427.10. Found: 392.1. (M−HCl+H)⁺.

Compound 65:2,6-Difluoro-N-[2′-methyl-5′-(1H-tetrazol-5-yl)-biphenyl-4-yl]-benzamide,sodium salt

¹H-NMR (CDCl₃) δ, 7.95-7.24 (m, 8H), 7.11-7.01 (m, 2H), 2.38 (s, 3H).ESMS clcd for C₂₁H₁₄F₂N₅NaO: 413.11. Found: 392.1. (M−Na+H)⁺.

Compound 44:3-Methyl-N-[2′-methyl-5′-(1H-tetrazol-5-yl)-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CDCl₃) δ, 8.58-7.31 (m, 10H), 2.52 (s, 3H), 2.37 (s, 3H). ESMSclcd for C₂₁H₁₈N₆O: 370.15. Found: 371.1. (M+H)⁺.

Compound 80:3-Methyl-N-[2′-methyl-5′-(1H-tetrazol-5-yl)-biphenyl-4-yl]-isonicotinamide,sodium salt

¹H-NMR (CD₃OD) δ, 9.06-7.31 (m, 10H), 2.63 (s, 3H), 2.32 (s, 3H). ESMSclcd for C₂₁H₁₇N₆NaO: 392.14. Found: 371.1. (M−Na+H)⁺.

Compound 28:3,5-Difluoro-N-[5′-(thiazol-2-yl)-2′-methyl-biphenyl-4-yl]-isonicotinamide

¹H-NMR (CDCl₃) δ, 8.45 (s, 2H), 7.93-7.31 (m, 9H), 2.43 (s, 3H). ESMSclcd for C₂₂H₁₅F₂N₃OS: 407.09. Found: 408.1. (M+H)⁺.

Other examples are listed in the following table:

# Chemical Name Structure ESMS 49 3-Methyl-N-[2′-methyl-5′-(1-methyl-1H-tetrazol-5-yl)- biphenyl-4-yl]-isonicotinamide

384.17 82 2,6-Difluoro-N-[2′-methyl- 5′-(1-methyl-1H-tetrazol-5-yl)-biphenyl-4-yl]-benzamide

405.14 83 2,6-Difluoro-N-[2′-methyl- 5′-(2-methyl-2H-tetrazol-5-yl)-biphenyl-4-yl]-benzamide

405.14

Example 2 Inhibition of IL-2 Production

Jurkat cells were placed in a 96 well plate (0.5 million cells per wellin 1% FBS medium) then a test compound of this invention was added atdifferent concentrations. After 10 minutes, the cells were activatedwith PHA (final concentration 2.5 μg/mL) and incubated for 20 hours at37° C. under CO₂. The final volume was 200 μL. Following incubation, thecells were centrifuged and the supernatants collected and stored at −70°C. prior to assaying for IL-2 production. A commercial ELISA kit (IL-2Eli-pair, Diaclone Research, Besancon, France) was used to detectproduction of IL-2, from which dose response curves were obtained. TheIC₅₀ value was calculated as the concentration at which 50% of maximumIL-2 production after stimulation was inhibited versus a non-stimulationcontrol.

Compound # IC₅₀ 1 2-4 nM 2 9 nM 3 1.6 nM 4 2 nM 5 1 nM 6 9 nM 7 8 nM 813 nM 9 7 nM 10 3 nM 11 3 nM 12 5 nM 13 6 nM 14 10 nM 15 2 nM 16 14 17 7nM 18 9 nM 19 19 nM 20 8 nM 21 6 nM 22 1 nM 23 5 nM 24 15 nM 25 3 nM 265 nM 27 4 nM 28 7 nM 29 9 nM 30 3 nM 31 6 nM 32 4 nM 33 4 nM 34 11 nM 352 nM 36 4 nM 37 11 nM 38 3 nM 39 13 nM 40 11 nM 41 18 nM 42 >1000 nM 4326 nM 44 >1000 nM 45 50 nM 47 59 nM 48 320 nM 49 12 nM 50 23 nM 51 11 nM52 35 nM 53 6 nM 54 12 nM 55 6 nM 56 2 nM 57 3 nM 58 2 nM 59 3 nM 60 13nM 61 6 nM 62 19 nM 63 >1000 nM 64 53 nM 65 >1000 nM 66 8 nM 67 27 nM 683 nM 69 5 nM 70 13 nM 71 6 nM 72 >1000 nM 73 >1000 nM 74 >1000 nM75 >1000 nM 76 7 nM 77 10 nM 78 9 nM 79 10 80 >1000 81 138 82 8 83 317

Inhibition of other cytokines, such as IL-4, IL-5, IL-13, GM-CSF, TNF-α,and INF-γ, can be tested in a similar manner using a commerciallyavailable ELISA kit for each cytokine.

Example 3 Patch Clamp Studies of Inhibition of I_(CRAC) Current in RBLCells, Jurkat Cells, and Primary T Cells

In general, a whole cell patch clamp method was used to examine theeffects of a compound of the invention on a channel that mediatesI_(crac). In such experiments, a baseline measurement was establishedfor a patched cell. Then a compound to be tested was perfused (orpuffed) to cells in the external solution and the effect of the compoundon I_(crac) was measured. A compound that modulates I_(crac) (e.g.,inhibits) is a compound that is useful in the invention for modulatingCRAC ion channel activity.

1) RBL Cells

Cells

Rat basophilic leukemia cells (RBL-2H3) were grown in DMEM mediasupplemented with 10% fetal bovine serum in an atmosphere of 95% air/5%CO₂. Cells were seeded on glass coverslips 1-3 days before use.

Recording Conditions

Membrane currents of individual cells were recorded using the whole-cellconfiguration of the patch clamp technique with an EPC10 (HEKAElectronik, Lambrecht, Germany). Electrodes (2-5 MΩ in resistance) werefashioned from borosilicate glass capillary tubes (Sutter Instruments,Novato, Ca). The recordings were done at room temperature.

Intracellular Pipette Solution

The intracellular pipette solution contained Cs-Glutamate 120 mM; CsCl20 mM; CsBAPTA 10 mM; CsHEPES 10 mM; NaCl 8 mM; MgCl₂ 1 mM; IP3 0.02 mM;pH=7.4 adjusted with CsOH. The solution was kept on ice and shieldedfrom light before the experiment was preformed.

Extracellular Solution

The extracellular solution contained NaCl 138 mM; NaHEPES, 10 mM; CsCl10 mM; CaCl₂ 10 mM; Glucose 5.5 mM; KCl 5.4 mM; KH₂PO₄ 0.4 mM;Na₂HPO₄.H₂O 0.3 mM at pH=7.4 adjusted with NaOH.

Compound Treatment

Each compound was diluted from a 10 mM stock in series using DMSO. Thefinal DMSO concentration was always kept at 0.1%.

Experimental Procedure

I_(CRAC) currents were monitored every 2 seconds using a 50 msecprotocol, where the voltage was ramped from −100 mV to +100 mV. Themembrane potential was held at 0 mV between the test ramps. In a typicalexperiment, the peak inward currents would develop within 50-100seconds. Once the I_(CRAC) currents were stabilized, the cells wereperfused with a test compound in the extracellular solution. At the endof an experiment, the remaining I_(CRAC) currents were then challengedwith a control compound (SKF96365, 10 μM) to ensure that the currentcould still be inhibited.

Data Analysis

The I_(CRAC) current level was determined by measuring the inwardcurrent amplitude at −80 mV of the voltage ramp in an off-line analysisusing MATLAB. The I_(CRAC) current inhibition for each concentration wascalculated using peak amplitude in the beginning of the experiment fromthe same cell. The IC₅₀ value and Hill coefficient for each compound wasestimated by fitting all the individual data points to a single Hillequation.

Results

The table below shows the concentration of compounds of the inventionwhich inhibits 50% of the I_(CRAC) current in RBL cells. As can be seenfrom the data in the table, representative compounds of the inventioninhibit I_(CRAC) current at concentration of 30 nM or less.

Compound Number IC₅₀ 1 20 nM 3 30 nM 4 90 nM 5 80 nM 7 70 nM 8 330 nM 9150 nM 10 40 nM 11 60 nM 12 20 nM 14 80 nM 15 70 nM 16 80 nM 17 40 nM 26160 nM 32 60 nM 36 130 nM 58 40 nM 70 390 mM SKF96365 4 μM

2) Jurkat Cells

Cells

Jurkat T cells were grown on glass coverslips, transferred to therecording chamber and kept in a standard modified Ringer's solution ofthe following composition: NaCl 145 mM, KCl 2.8 mM, CsCl 10 mM, CaCl₂ 10mM, MgCl₂ 2 mM, glucose 10 mM, HEPES.NaOH 10 mM, pH 7.2.

Extracellular Solution

The external solution contained 10 mM CaNaR, 11.5 mM glucose and a testcompound at various concentrations.

Intracellular Pipette Solution

The standard intracellular pipette solution contained: Cs-glutamate 145mM, NaCl 8 mM, MgCl₂ 1 mM, ATP 0.5 mM, GTP 0.3 mM, pH 7.2 adjusted withCsOH. The solution was supplemented with a mixture of 10 mM Cs-BAPTA and4.3-5.3 mM CaCl₂ to buffer [Ca²⁺]i to resting levels of 100-150 nM.

Patch-Clamp Recordings

Patch-clamp experiments were performed in the tight-seal whole-cellconfiguration at 21-25° C. High-resolution current recordings wereacquired by a computer-based patch-clamp amplifier system (EPC-9, HEKA,Lambrecht, Germany). Sylgard®-coated patch pipettes had resistancesbetween 2-4 MΩ after filling with the standard intracellular solution.Immediately following establishment of the whole-cell configuration,voltage ramps of 50 ms duration spanning the voltage range of −100 to+100 mV were delivered from a holding potential of 0 mV at a rate of 0.5Hz over a period of 300 to 400 seconds. All voltages were corrected fora liquid junction potential of 10 mV between external and internalsolutions. Currents were filtered at 2.3 kHz and digitized at 100 μsintervals. Capacitive currents and series resistance were determined andcorrected before each voltage ramp using the automatic capacitancecompensation of the EPC-9.

Data Analysis

The very first ramps before activation of I_(CRAC) (usually 1 to 3) weredigitally filtered at 2 kHz, pooled and used for leak-subtraction of allsubsequent current records. The low-resolution temporal development ofinward currents was extracted from the leak-corrected individual rampcurrent records by measuring the current amplitude at −80 mV or avoltage of choice.

Compound 1 was determined to be a strong inhibitor of I_(CRAC) in humanJurkat T cells.

3) Primary T Cells

Preparation of Primary T Cells

Primary T cells are obtained from human whole blood samples by adding100 μL of RosetteSep® human T cell enrichment cocktail to 2 mL of wholeblood. The mixture is incubated for 20 minutes at room temperature, thendiluted with an equal volume of PBS containing 2% FBS. The mixture islayered on top of RosetteSep® DM-L density medium and then centrifugedfor 20 minutes at 1200 g at room temperature. The enriched T cells arerecovered from the plasma/density medium interface, then washed with PBScontaining 2% FBS twice, and used in patch clamp experiments followingthe procedure described for RBL cells.

Example 4 Inhibition of Multiple Cytokines in Primary Human PBMCs

Peripheral blood mononuclear cells (PBMCs) are stimulated withphytohaemagglutinin (PHA) in the presence of varying concentrations ofcompounds of the invention or cyclosporine A (CsA), a known inhibitor ofcytokine production. Cytokine production is measured using commerciallyavailable human ELISA assay kits (from Cell Science, Inc.) following themanufacturers instructions.

The compounds of the invention are expected to be potent inhibitors ofIL-2, IL-4, IL-5, IL-13, GM-CSF, INF-γ and TNF-α in primary human PBMcells. In addition, compounds of the invention are not expected toinhibit the anti-inflammatory cytokine, IL-10.

Example 5 Compounds of the Invention are Potent Inhibitors ofDegranulation in RBL Cells

Procedure:

The day before the assay was performed, RBL cells, that had been grownto confluence in a 96 well plate, were incubated at 37° C. for at least2 hours. The medium was replaced in each well with 100 μL of freshmedium containing 2 μLg/mL of anti-DNP IgE.

On the following day, the cells were washed once with PRS (2.6 mMglucose and 0.1% BSA) and 160 μL of PRS was added to each well. A testcompound was added to a well in a 20 μL solution at 10× of the desiredconcentration and incubated for 20 to 40 minutes at 37° C. 20 μL of 10×mouse anti-IgE (10 μL/mL) was added. Maximum degranulation occurredbetween 15 to 40 minutes after addition of anti-IgE.

Results:

The table below shows the concentration of compounds of the inventionwhich inhibits 50% of degranulation in RBL cells. As can be seen fromthe data in the table below, representative compounds of the inventioninhibit degranulation at concentration of 4.5 μM or less. SKF96365 wasused as a positive control.

IC₅₀ Compound 1 2.52 μM Compound 2 4.85 μM Compound 3 4.5 μM Compound 41.45 μM Compound 5 1.1 μM Compound 6 5.01 μM Compound 8 0.52 μM Compound9 1.41 μM Compound 12 0.48 μM SKF96365 >20 μM

Example 6 Compounds of the Invention are Potent Inhibitors of Chemotaxisin T Cells

T-Cell Isolation:

Twenty ml aliquots of heparinized whole blood (2 pig, 1 human) weresubjected to density gradient centrifugation on Ficoll Hypaque. Thebuffy coat layers representing peripheral blood mononuclear cells(PBMCs) containing lymphocytes and monocytes were washed once,resuspended in 12 ml of incomplete RPMI 1640 and then placed ingelatin-coated T75 culture flasks for 1 hr at 37° C. The non-adherentcells, representing peripheral blood lymphocytes (PBLs) depleted ofmonocytes, were resuspended in complete RPMI media and placed in looselypacked activated nylon wool columns that had been equilibrated with warmmedia. After 1 hr at 37° C., the non-adherent T cell populations wereeluted by washing of the columns with additional media. The T cellpreparations were centrifuged, resuspended in 5 ml of incomplete RPMI,and counted using a hemocytometer.

Cell Migration Assay:

Aliquots of each T cell preparation were labeled with Calcien AM(TefLabs) and suspended at a concentration of 2.4×10⁶/ml inHEPES-buffered Hank's Balanced Salt Solution containing 1.83 mM CaCl₂and 0.8 mM MgCl₂, pH 7.4 (HHBSS). An equal volume of HHBSS containing 0,20 nM, 200 nM or 2000 nM of compound 1 or 20 nM EDTA was then added andthe cells incubated for 30 min at 37° C. Fifty μl aliquots of the cellsuspensions (60,000 cells) were placed on the membrane (pore size 5 μm)of a Neuroprobe ChemoTx 96 well chemotaxis unit that had been affixedover wells containing 10 ng/ml MIP-1α in HHBSS. The T cells were allowedto migrate for 2 hr at 37° C., after which the apical surface of themembrane was wiped clean of cells. The chemotaxis units were then placedin a CytoFlour 4000 (PerSeptive BioSystems) and the fluorescence of eachwell measured (excitation and emission wavelengths of 450 and 530 nm,respectively). The number of migrating cells in each well was determinedfrom a standard curve generated from measuring the fluorescence ofserial two-fold dilutions of the labeled cells placed in the lower wellsof the chemotaxis unit prior to affixing the membrane.

Results:

Compound 1 is inhibitory to the chemotactic response of porcine T cellsto 10 ng/ml MIP-1α (IC₅₀ values of ˜5 nM) and in human T cells to 100ng/ml MIP-1α (see FIG. 1). The data represent the averages oftriplicates. EDTA was used as a control compound in this assay (data notshown).

All publications, patent applications, patents, and other documentscited herein are incorporated by reference in their entirety. In case ofconflict, the present specification, including definitions, willcontrol. In addition, the materials, methods, and examples areillustrative only and not intended to be limiting in any way.

1. A compound represented by formula (I):

or a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof, wherein: L is a linker selected from the group consisting of acovalent bond, —NRCH₂—, —CH₂NR—, —C(O)—, —NR—C(O)—, —C(O)—NR—, —OC(O)—,—C(O)O—, —C(S)—, —NR—C(S)—, and —C(S)—NR—; X₁ and X₃ are each,independently, CH or N; X₂ is CH, CR₁₀ or N; each Z is independentlyselected from the group consisting of a lower alkyl, a lower haloalkyl,a halo, a lower alkoxy, a lower alkyl sulfanyl, cyano, nitro, and lowerhaloalkoxy; R, for each occurrence is independently selected from —H, analkyl, —C(O)R₅, and —C(O)OR₅; R₁ and R₂ are each, independently, a halo,a haloalkyl, a lower alkyl, a lower alkoxy, or a haloalkoxy; R₃ is analkyl, a haloalkyl, a halo, a haloalkoxy, —OR₅, —SR₅, or —NR₆R₇; R₁₈ isa halo, cyano, nitro, —C(O)R₅, —C(O)OR₅, —C(O)SR₅, —C(O)NR₆R₇, —C(S)R₅,—C(S)OR₅, —C(S)SR₅, —C(S)NR₆R₇, —C(NR₈)R₅, —C(NR₈)OR₅, —C(NR₈)SR₅,—C(NR₈)NR₆R₇, —S(O)_(p)R₅, —S(O)_(p)NR₅, —S(O)_(p)OR₅, —P(O)(OR₅)₂,—OP(O)(OR₅)₂, —P(O)(R₅)₂, a five or six membered optionally substitutedheterocycloalkyl, a five or six membered optionally substitutedheterocyclyl, or a five or six membered optionally substitutedheteroaryl; R₅, for each occurrence, is independently, H, an optionallysubstituted alkyl, an optionally substituted alkenyl, an optionallysubstituted alkynyl, an optionally substituted cycloalkyl, an optionallysubstituted cycloalkenyl, an optionally substituted heterocyclyl, anoptionally substituted aryl, an optionally substituted heteroaryl, anoptionally substituted aralkyl, or an optionally substitutedheteraralkyl; R₆ and R₇, for each occurrence are, independently, H, anoptionally substituted alkyl, an optionally substituted alkenyl, anoptionally substituted alkynyl, an optionally substituted cycloalkyl, anoptionally substituted cycloalkenyl, an optionally substitutedheterocyclyl, an optionally substituted aryl, an optionally substitutedheteroaryl, an optionally substituted aralkyl, or an optionallysubstituted heteraralkyl; or R₆ and R₇ taken together with the nitrogento which they are attached are an optionally substituted heterocyclyl oroptionally substituted heteroaryl; R₈, for each occurrence, isindependently —H, a halo, an alkyl, —OR₅, —NR₆R₇, —C(O)R₅, —C(O)OR₅, or—C(O)NR₆R₇; R₁₀ is a lower alkyl, a lower alkoxy, a halo, a lowerhaloalkyl, a lower haloalkoxy, a cyano, nitro, —C(O)R₅, —C(O)OR₅,—C(O)SR₅, —C(O)NR₆R₇, —C(S)R₅, —C(S)OR₅, —C(S)SR₅, —C(S)NR₆R₇,—C(NR₈)R₅, —C(NR₈)OR₅, —C(NR₈)SR₅, —C(NR₈)NR₆R₇, —S(O)_(p)R₅,—P(O)(OR₅)₂, —OP(O)(OR₅)₂, or —P(O)(R₅)₂; n is zero or an integer from 1to 4; and p, for each occurrence, is independently 1 or
 2. 2. Thecompound of claim 1, wherein L is —NR—C(O)—. 3-7. (canceled)
 8. Thecompound of claim 1, wherein the compound is represented by formula(II):

or a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof, wherein: R₄ is a halo, cyano, nitro, —C(O)R₅, —C(O)OR₅,—C(O)SR₅, —C(O)NR₆R₇, —C(S)R₅, —C(S)OR₅, —C(S)SR₅, —C(S)NR₆R₇,—C(NR₈)R₅, —C(NR₈)OR₅, —C(NR₈)SR₅, —C(NR₈)NR₆R₇, —S(O)_(p)R₅,—S(O)_(p)NR₅, —S(O)_(p)OR₅, —P(O)(OR₅)₂, —OP(O)(OR₅)₂, —P(O)(R₅)₂, or anester, amide or carboxylic acid bioisostere. 9-10. (canceled)
 11. Thecompound of claim 8, wherein L is —NHC(O)—. 12-19. (canceled)
 20. Thecompound of claim 11, wherein the compound is represented by structuralformula (IV):

or a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof. 21-27. (canceled)
 28. The compound of claim 11, wherein thecompound is represented by structural formula (V):

or a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.
 29. The compound of claim 28, wherein R₁ and R₂ are each,independently, a halo.
 30. The compound of claim 29, wherein R₃ is alower alkyl, a lower alkoxy, a lower alkyl sulfanyl, a lower alkylamino,a lower dialkylamino, or a halo.
 31. The compound of claim 30, R₄ is abioisostere of an ester, amide, or carboxylic acid.
 32. The compound ofclaim 31, wherein R₄ is a 5-membered heteroaryl.
 33. The compound ofclaim 32, wherein R₄ is an optionally substituted oxazolyl, anoptionally substituted thiazolyl, an optionally substituted1H-tetrazolyl, an optionally substituted 1H-imidazolyl, an optionallysubstituted [1,2,4]oxadiazolyl, or an optionally substituted4H-[1,2,4]-triazolyl.
 34. The compound of claim 30, wherein R₄ is ahalo, —C(O)R₉, —S(O)_(p)R₁₁, —S(O)_(p)NR₅, —S(O)_(p)OR₅, —P(O)(OR₁₂)₂,or —P(O)(R₁₁)₂, wherein: R₉ is a lower alkoxy, lower alkyl sulfanyl, oran alkoxyalkoxy; R₁₁, for each occurrence, is independently, a loweralkyl; and R₁₂, for each occurrence, is independently, H or a loweralkyl.
 35. The compound of claim 30, wherein R₁ and R₂ are each a fluorogroup.
 36. The compound of claim 11, wherein the compound is representedby structural formula (VI):

or a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof. 37-44. (canceled)
 45. A compound represented by formula (VII):

or a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof, wherein: L is a linker selected from the group consisting of acovalent bond, —NRCH₂—, —CH₂NR—, —C(O)—, —NR—C(O)—, —C(O)—NR—, —OC(O)—,—C(O)O—, —C(S)—, —NR—C(S)—, and —C(S)—NR—; X₁ is CH or N; each Z isindependently selected from the group consisting of a lower alkyl, alower haloalkyl, a halo, a lower alkoxy, a lower alkyl sulfanyl, cyano,nitro, or lower haloalkoxy; R, for each occurrence is independentlyselected from —H, an alkyl, —C(O)R₅, and —C(O)OR₅; R₁ is a halo, ahaloalkyl, a lower alkyl, a lower alkoxy, or a haloalkoxy; R₃ is analkyl, a haloalkyl, a halo, a haloalkoxy, —OR₅, —SR₅, or —NR₆R₇; R₁₈ isa halo, cyano, nitro, —C(O)R₅, —C(O)OR₅, —C(O)SR₅, —C(O)NR₆R₇, —C(S)R₅,—C(S)OR₅, —C(S)SR₅, —C(S)NR₆R₇, —C(NR₈)R₅, —C(NR₈)OR₅, —C(NR₈)SR₅,—C(NR₈)NR₆R₇, —S(O)_(p)R₅, —S(O)_(p)NR₅, —S(O)_(p)OR₅, —P(O)(OR₅)₂,—OP(O)(OR₅)₂, —P(O)(R₅)₂, a five or six membered optionally substitutedheterocycloalkyl, a five or six membered optionally substitutedheterocyclyl, or a five or six membered optionally substitutedheteroaryl; R₅, for each occurrence, is independently, H, an optionallysubstituted alkyl, an optionally substituted alkenyl, an optionallysubstituted alkynyl, an optionally substituted cycloalkyl, an optionallysubstituted cycloalkenyl, an optionally substituted heterocyclyl, anoptionally substituted aryl, an optionally substituted heteroaryl, anoptionally substituted aralkyl, or an optionally substitutedheteraralkyl; R₆ and R₇, for each occurrence are, independently, H, anoptionally substituted alkyl, an optionally substituted alkenyl, anoptionally substituted alkynyl, an optionally substituted cycloalkyl, anoptionally substituted cycloalkenyl, an optionally substitutedheterocyclyl, an optionally substituted aryl, an optionally substitutedheteroaryl, an optionally substituted aralkyl, or an optionallysubstituted heteraralkyl; or R₆ and R₇ taken together with the nitrogento which they are attached are an optionally substituted heterocyclyl oroptionally substituted heteroaryl; R₈, for each occurrence, isindependently —H, a halo, an alkyl, —OR₅, —NR₆R₇, —C(O)R₅, —C(O)OR₅, or—C(O)NR₆R₇; n is zero or an integer from 1 to 4; and p, for eachoccurrence, is independently 1 or
 2. 46. The compound of claim 45,wherein L is —NR—C(O)—. 47-51. (canceled)
 52. The compound of claim 45,wherein the compound is represented by formula (VI):

or a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof, wherein: R₄ is a halo, cyano, nitro, —C(O)R₅, —C(O)OR₅,—C(O)SR₅, —C(O)NR₆R₇, —C(S)R₅, —C(S)OR₅, —C(S)SR₅, —C(S)NR₆R₇,—C(NR₈)R₅, —C(NR₈)OR₅, —C(NR₈)SR₅, —C(NR₈)NR₆R₇, —S(O)_(p)R₅,—S(O)_(p)NR₅, —S(O)_(p)OR₅, —P(O)(OR₅)₂, —OP(O)(OR₅)₂, —P(O)(R₅)₂, or anester, amide or carboxylic acid bioisostere. 53-54. (canceled)
 55. Thecompound of claim 52, wherein L is —NHC(O)—.
 56. The compound of claim55, wherein the compound is represented by formula (IX):

or a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof. 57-115. (canceled)
 116. A pharmaceutical composition,comprising a pharmaceutically acceptable carrier and a compound of claim1 and optionally further comprising one or more additional therapeuticagents. 117-119. (canceled)
 120. A pharmaceutical composition,comprising a pharmaceutically acceptable carrier and a compound of claim45 and optionally further comprising one or more additional therapeuticagents.